Extended Data Fig. 6 | Peroxisomes and the ether lipid biosynthesis
pathway contribute to ferroptosis in liver, endometrial and kidney
cancers. a, Fluorescent imaging analysis of peroxisome abundances, reported
by PTS1–emGFP signal, in HuH-7 cells expressing the indicated sgRNAs. Scale
bar, 25 μm. Representative images from experiment performed once, and each
condition has three biological replicates. b, Violin plots showing the
quantitation of peroxisomes in HuH-7 cells expressing indicated sgRNAs. HuH-
7 sgNC, n = 2 50; PEX3 sg1, n = 2 31; PEX3 sg2, n = 2 56; PEX10 sg1, n = 600; PEX10
sg2, n = 455; PEX12 sg1, n = 295; PEX12 sg2, n = 278; AGPS sg1, n = 321; AGPS sg2,
n = 30 4; FA R 1 sg1, n = 411. Lines in violin plots indicate median and quartiles.
c, Immunoblot analysis of AGPS protein levels in HuH-7 cells expressing
negative control (sgNC) or AGPS-targeting sgRNAs. β-Actin was used as a
loading control. Plot of experiment performed once. d, Viability curves for
HuH-7 cells expressing the indicated sgRNAs treated with indicated
concentrations of ML210 or RSL3 for 48 h. n = 4 biologically independent
samples. Representative results from experiment performed twice. e, Viability
curves for SNU-685 cells expressing the indicated sgRNAs treated with
indicated concentrations of ML210 or RSL3 for 48 h. n = 4 biologically
independent samples. Representative results from experiment performed
once. f, Volcano plots showing the lipidomic analysis results^15 comparing 49
pairs of ccRCC tumour and adjacent normal kidney tissues. n = 49 tumour
samples, n = 49 normal samples. Two-tailed Student’s t-test. Multiple-testing
adjustment was performed using the Benjamini–Hochberg method.
See Supplementary Information for uncropped immunoblot images. For
viability curves, data are mean ± s.d.