def _plot_with_rpy2(self, regions, filename):
from rpy2 import robjects
import rpy2.robjects.lib.ggplot2 as ggplot2
from rpy2.robjects.lib import grid
from rpy2.robjects.packages import importr
grdevices = importr('grDevices')
base = importr('base')
grdevices.pdf(file=filename + '.pdf')
t = [x for x in range(-self.num_bins, self.num_bins + 1)]
for region in regions[:self.num_regs]:
if not np.any(region.weighted):
logger.warning(
"Warning: No data for region located on bin " + str(region.bin) + ". Not plotting this one.")
continue
middle = (len(region.weighted[0]) - 1) / 2
if middle < self.num_bins:
logger.error("Warning: There are less bins calculated for regions than you want to plot.")
sys.exit(1)
d = {'map': robjects.StrVector(
[str(m) for sublist in [[x] * len(t) for x in range(len(region.weighted))] for m in sublist]),
't': robjects.FloatVector(t * len(region.weighted)),
'e': robjects.FloatVector([i for sublist in region.weighted for i in
sublist[middle - self.num_bins:middle + self.num_bins + 1]]),
'p': robjects.FloatVector([-np.log10(x) for sublist in region.pvalues for x in
sublist[middle - self.num_bins:middle + self.num_bins + 1]]),
'c': robjects.FloatVector([-np.log10(x) for sublist in region.corrected_pvalues for x in
sublist[middle - self.num_bins:middle + self.num_bins + 1]])}
dataf = robjects.DataFrame(d)
gp = ggplot2.ggplot(dataf) # first yellow second red
p1 = gp + ggplot2.geom_line(mapping=ggplot2.aes_string(x='t', y='e', group='map', colour='map'),
alpha=0.8) + ggplot2.scale_y_continuous(trans='log2') + ggplot2.ggtitle(
"\n".join(wrap("Bin " + str(region.bin) + " : " + str(region.positions)))) + ggplot2.labs(
y="log Intensity") + ggplot2.theme_classic() + ggplot2.theme(
**{'axis.title.x': ggplot2.element_blank(), 'axis.text.y': ggplot2.element_text(angle=45),
'axis.text.x': ggplot2.element_blank(),
'legend.position': 'none'}) + ggplot2.scale_colour_brewer(palette="Set1")
p2 = gp + ggplot2.geom_line(mapping=ggplot2.aes_string(x='t', y='p', group='map', colour='map'),
alpha=0.8) + ggplot2.labs(
y="-log10(p-value)") + ggplot2.theme_classic() + ggplot2.theme(
**{'axis.title.x': ggplot2.element_blank(), 'axis.text.x': ggplot2.element_blank(),
'legend.position': 'none'}) + ggplot2.scale_colour_brewer(palette="Set1")
p3 = gp + ggplot2.geom_line(mapping=ggplot2.aes_string(x='t', y='c', group='map', colour='map'),
alpha=0.8) + ggplot2.labs(y="-log10(q-value)",
x='bins (' + str(self.bin_res) + ' bp each)') + \
ggplot2.geom_hline(mapping=ggplot2.aes_string(yintercept=str(-np.log10(self.threshold))),
colour='black', alpha=0.8, linetype='dashed') + ggplot2.theme_classic() + \
ggplot2.theme(**{'legend.position': 'none'}) + ggplot2.scale_colour_brewer(palette="Set1")
g1 = ggplot2.ggplot2.ggplotGrob(p1)
g2 = ggplot2.ggplot2.ggplotGrob(p2)
g3 = ggplot2.ggplot2.ggplotGrob(p3)
robjects.globalenv["g"] = base.rbind(g1, g2, g3, size='first')
robjects.r("grid::grid.draw(g)")
grid.newpage()
logger.debug('Plotted region ' + str(region.bin))
grdevices.dev_off()
def generate_histogram(subgroups_to_sses_to_n_count, tname, file_name):
columns_to_data = {'subgroup': [], tname: [], 'count': []}
max_count = 0
for subgroup, sses_to_n_count in subgroups_to_sses_to_n_count.items():
for ss, n_count in sses_to_n_count.items():
columns_to_data['subgroup'].append(subgroup)
columns_to_data[tname].append(ss)
columns_to_data['count'].append(n_count)
if n_count > max_count:
max_count = n_count
r_columns_to_data = {
'subgroup':
ro.FactorVector(columns_to_data['subgroup'],
levels=ro.StrVector(
_sort_subgroup(set(columns_to_data['subgroup'])))),
tname:
ro.StrVector(columns_to_data[tname]),
'count':
ro.IntVector(columns_to_data['count'])
}
df = ro.DataFrame(r_columns_to_data)
max_count = int(max_count / 1000 * 1000 + 1000)
histogram_file_path = os.path.join(OUTPUT_PATH, file_name)
logging.debug(
str.format("The Data Frame for file {}: \n{}", histogram_file_path,
df))
grdevices.png(file=histogram_file_path, width=1200, height=800)
gp = ggplot2.ggplot(df)
pp = gp + \
ggplot2.aes_string(x='subgroup', y='count', fill=tname) + \
ggplot2.geom_bar(position="dodge",width=0.8, stat="identity") + \
ggplot2.theme_bw() + \
ggplot2.theme_classic() + \
ggplot2.theme(**{'legend.title': ggplot2.element_blank()}) + \
ggplot2.theme(**{'legend.text': ggplot2.element_text(size=40)}) + \
ggplot2.theme(**{'axis.text.x': ggplot2.element_text(size=40,angle=45)}) + \
ggplot2.theme(**{'axis.text.y': ggplot2.element_text(size=40)}) + \
ggplot2.scale_y_continuous(expand=ro.IntVector([0, 0]),
limits=ro.IntVector([0, max_count])) + \
ggplot2.geom_text(ggplot2.aes_string(label='count'), size=6, angle=35, hjust=-0.1,
position=ggplot2.position_dodge(width=0.8),
vjust=-0.2)
pp.plot()
logging.info(str.format("Output step3 file {}", histogram_file_path))
grdevices.dev_off()
def generate_step3_5_lrr_acc20_line_chart(subgroups_to_lrrs_acc20mean,
prefix=''):
pandas2ri.activate()
subgroups_to_lrr_count = {}
columns_to_data = {'subgroup': [], 'pos': [], 'acc20': []}
for subgroup, (acc20means,
acc20_count) in subgroups_to_lrrs_acc20mean.items():
subgroups_to_lrr_count[subgroup] = acc20_count
for index, acc20mean in enumerate(acc20means):
columns_to_data['subgroup'].append(subgroup)
columns_to_data['pos'].append(index + 1)
columns_to_data['acc20'].append(acc20mean)
# Write the count of LRRs for each subgroup to file
with open(os.path.join(OUTPUT_PATH, prefix + "step3_5_lrr_count.txt"),
'w') as f:
for subgroup, lrr_count in subgroups_to_lrr_count.items():
f.write(str.format("{}: {}\n", subgroup, lrr_count))
# Generate the line chart file
r_columns_to_data = {
'subgroup': ro.StrVector(columns_to_data['subgroup']),
'pos': ro.IntVector(columns_to_data['pos']),
'acc20': ro.FloatVector(columns_to_data['acc20'])
}
df = ro.DataFrame(r_columns_to_data)
line_chart_file_path = os.path.join(OUTPUT_PATH,
prefix + "step3_5_lrr_acc20_line.png")
logging.debug(
str.format("The Data Frame for file {}: \n{}", line_chart_file_path,
df))
grdevices.png(file=line_chart_file_path, width=1024, height=512)
gp = ggplot2.ggplot(df)
pp = gp + \
ggplot2.theme_bw() + \
ggplot2.theme_classic() + \
ggplot2.theme(**{'axis.text.x': ggplot2.element_text(size=35)}) + \
ggplot2.theme(**{'axis.text.y': ggplot2.element_text(size=35)}) + \
ggplot2.aes_string(x='pos', y='acc20', group='subgroup', colour='subgroup') + \
ggplot2.geom_point(size=4, shape=20) + \
ggplot2.geom_line(size=3) + \
ggplot2.theme(**{'legend.title': ggplot2.element_blank()}) + \
ggplot2.theme(**{'legend.text': ggplot2.element_text(size=20)}) + \
ggplot2.scale_x_continuous(breaks=ro.IntVector(range(1, 25)), labels=ro.StrVector(list('LxxLxLxxNxLsGxIPxxLxxLxx')))
pp.plot()
logging.info(str.format("Output step3 file {}", line_chart_file_path))
grdevices.dev_off()
def generate_step3_9_n_count_histogram(place_type_pos_type_to_count,
file_name):
columns_to_data = {'place': [], 'pos': [], 'count': []}
max_count = 0
for place_pos_type, n_count in place_type_pos_type_to_count.items():
place_type, pos_type = place_pos_type.split('_')
columns_to_data['place'].append(place_type)
columns_to_data['pos'].append(pos_type)
columns_to_data['count'].append(n_count)
if n_count > max_count:
max_count = n_count
r_columns_to_data = {
'place': ro.StrVector(columns_to_data['place']),
'pos': ro.StrVector(columns_to_data['pos']),
'count': ro.IntVector(columns_to_data['count'])
}
df = ro.DataFrame(r_columns_to_data)
if max_count > 1000:
max_count = int(max_count / 1000 * 1000 + 1000)
else:
max_count = int(max_count / 100 * 100 + 100)
histogram_file_path = os.path.join(OUTPUT_PATH, file_name)
logging.debug(
str.format("The Data Frame for file {}: \n{}", histogram_file_path,
df))
grdevices.png(file=histogram_file_path, width=1024, height=512)
gp = ggplot2.ggplot(df)
pp = gp + \
ggplot2.aes_string(x='pos', y='count', fill='place') + \
ggplot2.geom_bar(position="dodge", stat="identity") + \
ggplot2.theme_bw() + \
ggplot2.theme_classic() + \
ggplot2.theme(**{'axis.text.x': ggplot2.element_text(size=35)}) + \
ggplot2.theme(**{'axis.text.y': ggplot2.element_text(size=35)}) + \
ggplot2.scale_y_continuous(expand=ro.IntVector([0, 0]),
limits=ro.IntVector([0, max_count])) + \
ggplot2.geom_text(ggplot2.aes_string(label='count'),
position=ggplot2.position_dodge(width=0.8), size=10, angle=35, hjust=-0.2,
vjust=-0.5)
pp.plot()
logging.info(str.format("Output step3 file {}", histogram_file_path))
grdevices.dev_off()
def _generate_step3_5_ss_acc20_line_chart(ts_to_acc20s, tname,
line_chart_file_path):
logging.debug(
str.format("Begin to generate {}, data {}", line_chart_file_path,
ts_to_acc20s))
ts_to_acc20mean = calc_acc20mean_by_types(ts_to_acc20s)
columns_to_data = {tname: [], 'site': [], 'acc20': []}
for ss, acc20means in ts_to_acc20mean.items():
for index, acc20mean in enumerate(acc20means):
columns_to_data[tname].append(ss)
columns_to_data['site'].append(index - 5)
columns_to_data['acc20'].append(acc20mean)
# Generate the line chart file
r_columns_to_data = {
tname: ro.StrVector(columns_to_data[tname]),
'site': ro.IntVector(columns_to_data['site']),
'acc20': ro.FloatVector(columns_to_data['acc20'])
}
df = ro.DataFrame(r_columns_to_data)
logging.debug(
str.format("The Data Frame for file {}: \n{}", line_chart_file_path,
df))
grdevices.png(file=line_chart_file_path, width=1024, height=512)
gp = ggplot2.ggplot(df)
pp = gp + \
ggplot2.theme_bw() + \
ggplot2.theme_classic() + \
ggplot2.theme(**{'axis.text.x': ggplot2.element_text(size=35)}) + \
ggplot2.theme(**{'axis.text.y': ggplot2.element_text(size=35)}) + \
ggplot2.aes_string(x='site', y='acc20', group=tname, colour=tname) + \
ggplot2.geom_point(size=4, shape=20) + \
ggplot2.geom_line(size=3) + \
ggplot2.theme(**{'legend.title': ggplot2.element_blank()}) + \
ggplot2.theme(**{'legend.text': ggplot2.element_text(size=20)}) + \
ggplot2.scale_x_continuous(breaks=ro.IntVector(list(range(-5, 6))),
labels=ro.StrVector(['-5', '-4', '-3', '-2', '-1', 'N', '1', '2', '3', '4', '5']))
pp.plot()
logging.info(str.format("Output step3 file {}", line_chart_file_path))
grdevices.dev_off()
def main():
'''
maine
'''
# Command Line Stuff...
myCommandLine = CommandLine()
outdir = myCommandLine.args['outDir']
group1 = myCommandLine.args['group1']
group2 = myCommandLine.args['group2']
batch = myCommandLine.args['batch']
matrix = myCommandLine.args['matrix']
prefix = myCommandLine.args['prefix']
formula = myCommandLine.args['formula']
print("running DESEQ2 %s" % prefix, file=sys.stderr)
# make the quant DF
quantDF = pd.read_table(matrix, header=0, sep='\t', index_col=0)
df = pandas2ri.py2ri(quantDF)
# import formula
formulaDF = pd.read_csv(formula,header=0, sep="\t",index_col=0)
sampleTable = pandas2ri.py2ri(formulaDF)
if "batch" in list(formulaDF):
design = Formula("~ batch + condition")
else:
design = Formula("~ condition")
# import DESeq2
from rpy2.robjects.packages import importr
import rpy2.robjects.lib.ggplot2 as ggplot2
methods = importr('methods')
deseq = importr('DESeq2')
grdevices = importr('grDevices')
qqman = importr('qqman')
### RUN DESEQ2 ###
R.assign('df', df)
R.assign('sampleTable', sampleTable)
R.assign('design',design)
R('dds <- DESeqDataSetFromMatrix(countData = df, colData = sampleTable, design = design)')
R('dds <- DESeq(dds)')
R('name <- grep("condition", resultsNames(dds), value=TRUE)')
###
###
# Get Results and shrinkage values
res = R('results(dds, name=name)')
resLFC = R('lfcShrink(dds, coef=name)')
vsd = R('vst(dds,blind=FALSE)')
resdf = robjects.r['as.data.frame'](res)
reslfc = robjects.r['as.data.frame'](resLFC)
dds = R('dds')
### Plotting section ###
# plot MA and PC stats for the user
plotMA = robjects.r['plotMA']
plotDisp = robjects.r['plotDispEsts']
plotPCA = robjects.r['plotPCA']
plotQQ = robjects.r['qq']
# get pca data
if "batch" in list(formulaDF):
pcaData = plotPCA(vsd, intgroup=robjects.StrVector(("condition", "batch")), returnData=robjects.r['T'])
percentVar = robjects.r['attr'](pcaData, "percentVar")
else:
print(vsd)
pcaData = plotPCA(vsd, intgroup="condition", returnData=robjects.r['T'])
percentVar = robjects.r['attr'](pcaData, "percentVar")
# arrange
data_folder = os.path.join(os.getcwd(), outdir)
qcOut = os.path.join(data_folder, "%s_QCplots_%s_v_%s.pdf" % (prefix,group1,group2))
grdevices.pdf(file=qcOut)
x = "PC1: %s" % int(percentVar[0]*100) + "%% variance"
y = "PC2: %s" % int(percentVar[1]*100) + "%% variance"
if "batch" in list(formulaDF):
pp = ggplot2.ggplot(pcaData) + \
ggplot2.aes_string(x="PC1", y="PC2", color="condition", shape="batch") + \
ggplot2.geom_point(size=3) + \
robjects.r['xlab'](x) + \
robjects.r['ylab'](y) + \
ggplot2.theme_classic() + \
ggplot2.coord_fixed()
else:
pp = ggplot2.ggplot(pcaData) + \
ggplot2.aes_string(x="PC1", y="PC2", color="condition") + \
ggplot2.geom_point(size=3) + \
robjects.r['xlab'](x) + \
robjects.r['ylab'](y) + \
ggplot2.theme_classic() + \
ggplot2.coord_fixed()
pp.plot()
plotMA(res, ylim=robjects.IntVector((-3,3)), main="MA-plot results")
plotMA(resLFC, ylim=robjects.IntVector((-3,3)), main="MA-plot LFCSrhinkage")
plotQQ(reslfc.rx2('pvalue'), main="LFCSrhinkage pvalue QQ")
hh = ggplot2.ggplot(resdf) + \
ggplot2.aes_string(x="pvalue") + \
ggplot2.geom_histogram() + \
ggplot2.theme_classic() + \
ggplot2.ggtitle("pvalue distribution")
hh.plot()
plotDisp(dds, main="Dispersion Estimates")
grdevices.dev_off()
data_folder = os.path.join(os.getcwd(), outdir)
lfcOut = os.path.join(data_folder, "%s_%s_v_%s_deseq2_results_shrinkage.tsv" % (prefix,group1,group2))
resOut = os.path.join(data_folder, "%s_%s_v_%s_deseq2_results.tsv" % (prefix,group1,group2))
robjects.r['write.table'](reslfc, file=lfcOut, quote=False, sep="\t")
robjects.r['write.table'](resdf, file=resOut, quote=False, sep="\t")
def main():
'''
maine
'''
# Command Line Stuff...
myCommandLine = CommandLine()
outdir = myCommandLine.args['outDir']
group1 = myCommandLine.args['group1']
group2 = myCommandLine.args['group2']
batch = myCommandLine.args['batch']
matrix = myCommandLine.args['matrix']
prefix = myCommandLine.args['prefix']
formula = myCommandLine.args['formula']
# make the quant DF
quantDF = pd.read_table(matrix, header=0, sep='\t', index_col=0)
df = pandas2ri.py2ri(quantDF)
#print(df.head())
# import formula
formulaDF = pd.read_csv(formula,header=0, sep="\t",index_col=0)
sampleTable = pandas2ri.py2ri(formulaDF)
if "batch" in list(formulaDF):
design = Formula("~ batch + condition")
else:
design = Formula("~ condition")
#print(sampleTable)
# import DESeq2
from rpy2.robjects.packages import importr
import rpy2.robjects.lib.ggplot2 as ggplot2
methods = importr('methods')
deseq = importr('DESeq2')
grdevices = importr('grDevices')
qqman = importr('qqman')
dds = deseq.DESeqDataSetFromMatrix(countData = df,
colData = sampleTable,
design = design)
dds = deseq.DESeq(dds)
cont = robjects.r["grep"]("condition",robjects.r['resultsNames'](dds),value=True)
#print(cont)
# get results; orient the results for groupA vs B
res = deseq.results(dds, name=cont)
# results with shrinkage
resLFC = deseq.lfcShrink(dds, coef=cont, type="apeglm")
resdf = robjects.r['as.data.frame'](res)
R.assign('res', res)
reslfc = robjects.r['as.data.frame'](resLFC)
# plot MA and PC stats for the user
plotMA = robjects.r['plotMA']
plotDisp = robjects.r['plotDispEsts']
plotPCA = robjects.r['plotPCA']
plotQQ = robjects.r['qq']
vsd = robjects.r['vst'](dds, blind=robjects.r['F'])
# get pca data
if "batch" in list(formulaDF):
pcaData = plotPCA(vsd, intgroup=robjects.StrVector(("condition", "batch")), returnData=robjects.r['T'])
percentVar = robjects.r['attr'](pcaData, "percentVar")
else:
print(vsd)
pcaData = plotPCA(vsd, intgroup="condition", returnData=robjects.r['T'])
percentVar = robjects.r['attr'](pcaData, "percentVar")
# arrange
grdevices.pdf(file="./%s/%s_QCplots_%s_v_%s.pdf" % (outdir,prefix,group1,group2))
x = "PC1: %s" % int(percentVar[0]*100) + "%% variance"
y = "PC2: %s" % int(percentVar[1]*100) + "%% variance"
if "batch" in list(formulaDF):
pp = ggplot2.ggplot(pcaData) + \
ggplot2.aes_string(x="PC1", y="PC2", color="condition", shape="batch") + \
ggplot2.geom_point(size=3) + \
robjects.r['xlab'](x) + \
robjects.r['ylab'](y) + \
ggplot2.theme_classic() + \
ggplot2.coord_fixed()
pp.plot()
else:
pp = ggplot2.ggplot(pcaData) + \
ggplot2.aes_string(x="PC1", y="PC2", color="condition") + \
ggplot2.geom_point(size=3) + \
robjects.r['xlab'](x) + \
robjects.r['ylab'](y) + \
ggplot2.theme_classic() + \
ggplot2.coord_fixed()
pp.plot()
plotMA(res, ylim=robjects.IntVector((-3,3)), main="MA-plot results")
#plotMA(res, main="MA-plot results")
plotMA(resLFC, ylim=robjects.IntVector((-3,3)), main="MA-plot LFCSrrhinkage")
#plotMA(resLFC, main="MA-plot LFCSrrhinkage")
plotQQ(resdf.rx2('pvalue'), main="pvalue QQ")
plotQQ(reslfc.rx2('pvalue'), main="LFCSrhinkage pvalue QQ")
hh = ggplot2.ggplot(resdf) + \
ggplot2.aes_string(x="pvalue") + \
ggplot2.geom_histogram() + \
ggplot2.theme_classic()
hh.plot()
plotDisp(dds, main="Dispersion Estimates")
grdevices.dev_off()
lfcOut = "./%s/%s_%s_v_%s_deseq2_results_shrinkage.tsv" % (outdir,prefix,group1,group2)
resOut = "./%s/%s_%s_v_%s_deseq2_results.tsv" % (outdir,prefix,group1,group2)
robjects.r['write.table'](reslfc, file=lfcOut, quote=False, sep="\t")
robjects.r['write.table'](resdf, file=resOut, quote=False, sep="\t")
def main():
'''
maine
'''
# Command Line Stuff...
myCommandLine = CommandLine()
outdir = "diffExpOut"
group1 = myCommandLine.args['group1']
group2 = myCommandLine.args['group2']
batch = myCommandLine.args['batch']
matrix = myCommandLine.args['matrix']
prefix = "flair_diffexp"
# make the quant DF
quantDF = pd.read_table(matrix, header=0, sep='\t')
quantDF = quantDF.set_index('ids')
df = pandas2ri.py2ri(quantDF)
# now make the formula
with open(matrix) as l:
header = next(l).rstrip().split()[1:]
formula = [[x, x.split("_")[1], x.split("_")[-1]] for x in header]
formulaDF = pd.DataFrame(formula)
formulaDF.columns = ['sampleName', 'condition', 'batch']
formulaDF = formulaDF.set_index('sampleName')
sampleTable = pandas2ri.py2ri(formulaDF)
design = Formula("~ batch + condition")
print(sampleTable)
# import DESeq2
from rpy2.robjects.packages import importr
import rpy2.robjects.lib.ggplot2 as ggplot2
methods = importr('methods')
deseq = importr('DESeq2')
grdevices = importr('grDevices')
qqman = importr('qqman')
dds = deseq.DESeqDataSetFromMatrix(countData=df,
colData=sampleTable,
design=design)
dds = deseq.DESeq(dds)
cont = robjects.r["grep"]("condition",
robjects.r['resultsNames'](dds),
value="TRUE")
# get results; orient the results for groupA vs B
res = deseq.results(dds, name=cont)
# results with shrinkage
resLFC = deseq.lfcShrink(dds, coef=cont, type="apeglm")
resdf = robjects.r['as.data.frame'](res)
R.assign('res', res)
R('write.table(res, file="testres.tsv", quote=FALSE, col.names=NA)')
reslfc = robjects.r['as.data.frame'](resLFC)
# plot MA and PC stats for the user
plotMA = robjects.r['plotMA']
plotDisp = robjects.r['plotDispEsts']
plotPCA = robjects.r['plotPCA']
plotQQ = robjects.r['qq']
vsd = robjects.r['vst'](dds, blind=robjects.r['F'])
# get pca data
pcaData = plotPCA(vsd,
intgroup=robjects.StrVector(("condition", "batch")),
returnData=robjects.r['T'])
percentVar = robjects.r['attr'](pcaData, "percentVar")
# arrange
grdevices.pdf(file="./%s/%s_%s_vs_%s_%s_cutoff_plots.pdf" %
(outdir, prefix, group1, group2, str(batch)))
x = "PC1: %s" % int(percentVar[0] * 100) + "%% variance"
y = "PC2: %s" % int(percentVar[1] * 100) + "%% variance"
pp = ggplot2.ggplot(pcaData) + \
ggplot2.aes_string(x="PC1", y="PC2", color="condition", shape="batch") + \
ggplot2.geom_point(size=3) + \
robjects.r['xlab'](x) + \
robjects.r['ylab'](y) + \
ggplot2.theme_classic() + \
ggplot2.coord_fixed()
pp.plot()
plotMA(res, ylim=robjects.IntVector((-3, 3)), main="MA-plot results")
#plotMA(res, main="MA-plot results")
plotMA(resLFC,
ylim=robjects.IntVector((-3, 3)),
main="MA-plot LFCSrrhinkage")
#plotMA(resLFC, main="MA-plot LFCSrrhinkage")
plotQQ(resdf.rx2('pvalue'), main="pvalue QQ")
plotQQ(reslfc.rx2('pvalue'), main="LFCSrhinkage pvalue QQ")
hh = ggplot2.ggplot(resdf) + \
ggplot2.aes_string(x="pvalue") + \
ggplot2.geom_histogram() + \
ggplot2.theme_classic()
hh.plot()
plotDisp(dds, main="Dispersion Estimates")
grdevices.dev_off()
lfcOut = "./%s/%s_%s_deseq2_results_LFC.tsv" % (outdir, prefix, str(batch))
resOut = "./%s/%s_%s_deseq2_results.tsv" % (outdir, prefix, str(batch))
robjects.r['write.table'](reslfc, file=lfcOut, quote=False, sep="\t")
robjects.r['write.table'](resdf, file=resOut, quote=False, sep="\t")
sys.exit(1)
reslsf = pandas2ri.ri2py(reslfc)
res = pandas2ri.ri2py(resdf)
reslsf.to_csv("./%s/%s_%s_deseq2_results_LFC.tsv" %
(outdir, prefix, str(batch)),
sep='\t')
res.to_csv("./%s/%s_%s_deseq2_results.tsv" % (outdir, prefix, str(batch)),
sep='\t')
def main():
'''
maine
'''
# Command Line Stuff...
myCommandLine = CommandLine()
workingdir = myCommandLine.args['workingdir']
outdir = myCommandLine.args['outdir']
group1 = myCommandLine.args['group1']
group2 = myCommandLine.args['group2']
batch = myCommandLine.args['batch']
files = myCommandLine.args['files']
prefix = myCommandLine.args['out_prefix']
sFilter = myCommandLine.args['filter']
makeDir(outdir)
files = checkSamples(files)
df = filesToDF(files, sFilter)
# DO DESEQ2
from rpy2 import robjects
from rpy2.robjects import r, pandas2ri, Formula
from rpy2.robjects.lib import grid
pandas2ri.activate()
# Compile data for data frame
data = list()
for f in files:
if group1 in f:
if batch in f:
data.append((f, group1, '1'))
else:
data.append((f, group1, '2'))
else:
if batch in f:
data.append((f, group2, '1'))
else:
data.append((f, group2, '2'))
# Make the Data Frame
pydf = pd.DataFrame(data)
pydf.columns = ['sampleName', 'condition', 'batch']
pydf = pydf.set_index('sampleName')
# Convert pandas to R data frame.
sampleTable = pandas2ri.py2ri(pydf)
# DESEQ2 part.
# Forumla
design = Formula("~ batch + condition")
# import DESeq2
from rpy2.robjects.packages import importr
import rpy2.robjects.lib.ggplot2 as ggplot2
methods = importr('methods')
deseq = importr('DESeq2')
grdevices = importr('grDevices')
qqman = importr('qqman')
# dds = deseq.DESeqDataSetFromHTSeqCount(sampleTable = sampleTable,
# directory = workingdir,
# design= design)
dds = deseq.DESeqDataSetFromMatrix(countData=df,
colData=sampleTable,
design=design)
dds = deseq.DESeq(dds)
# get results; orient the results for groupA vs B
res = deseq.results(dds,
contrast=robjects.StrVector(
("condition", group2, group1)))
# results with shrinkage
resLFC = deseq.lfcShrink(dds,
coef="condition_%s_vs_%s" % (group2, group1),
type="apeglm")
resdf = robjects.r['as.data.frame'](res)
reslfc = robjects.r['as.data.frame'](resLFC)
# plot MA and PC stats for the user
plotMA = robjects.r['plotMA']
plotDisp = robjects.r['plotDispEsts']
plotPCA = robjects.r['plotPCA']
plotQQ = robjects.r['qq']
vsd = robjects.r['vst'](dds, blind=robjects.r['F'])
# get pca data
pcaData = plotPCA(vsd,
intgroup=robjects.StrVector(("condition", "batch")),
returnData=robjects.r['T'])
percentVar = robjects.r['attr'](pcaData, "percentVar")
# arrange
grdevices.pdf(file="./%s/%s_%s_vs_%s_%s_%s_cutoff_plots.pdf" %
(outdir, prefix, group1, group2, str(batch), sFilter))
x = "PC1: %s" % int(percentVar[0] * 100) + "%% variance"
y = "PC2: %s" % int(percentVar[1] * 100) + "%% variance"
pp = ggplot2.ggplot(pcaData) + \
ggplot2.aes_string(x="PC1", y="PC2", color="condition", shape="batch") + \
ggplot2.geom_point(size=3) + \
robjects.r['xlab'](x) + \
robjects.r['ylab'](y) + \
ggplot2.theme_classic() + \
ggplot2.coord_fixed()
pp.plot()
plotMA(res, ylim=robjects.IntVector((-3, 3)), main="MA-plot results")
#plotMA(res, main="MA-plot results")
plotMA(resLFC,
ylim=robjects.IntVector((-3, 3)),
main="MA-plot LFCSrrhinkage")
#plotMA(resLFC, main="MA-plot LFCSrrhinkage")
plotQQ(resdf.rx2('pvalue'), main="pvalue QQ")
plotQQ(reslfc.rx2('pvalue'), main="LFCSrhinkage pvalue QQ")
hh = ggplot2.ggplot(resdf) + \
ggplot2.aes_string(x="pvalue") + \
ggplot2.geom_histogram() + \
ggplot2.theme_classic()
hh.plot()
plotDisp(dds, main="Dispersion Estimates")
grdevices.dev_off()
reslsf = pandas2ri.ri2py(reslfc)
res = pandas2ri.ri2py(resdf)
reslsf.to_csv("./%s/%s_%s_vs_%s_%s_deseq2_results_LFC.tsv" %
(outdir, prefix, group1, group2, str(batch)),
sep='\t')
reslsf.to_csv("./%s/%s_%s_vs_%s_%s_deseq2_results.tsv" %
(outdir, prefix, group1, group2, str(batch)),
sep='\t')
