a, HIF-2α expression by immunohistochemistry (IHC)
in sensitive (green) and resistant (red) tumors. Scale bars = 50 µM.
(Images representative of quantitative data shown in 3b.) b,
Quantification of HIF-2α-positive cells as determined by IHC in
sensitive, intermediate, and resistant tumors from all 22 tumorgraft lines
(Sensitive: n = 10; Intermediate: n = 5;
Resistant: n = 7). c, Western blot analysis of
sensitive (green) and resistant (red) tumorgraft lines. XP164 lysate loaded
twice as a reference for comparison between the two membranes. d,
qRT-PCR of EPAS1 (HIF-2α) expression in sensitive (n =
11) versus resistant (n = 26) vehicle-treated tumorgrafts.
e, Candidate genes from RNAseq analysis differentially
expressed in sensitive and resistant tumors. b: An ANOVA test was
used to determine if sensitive tumors were different from intermediate or
resistant. Bar chart depicts the mean with the error bar representing s.e.m.
d, e: Tests completed using a mixed model analysis with
compound symmetrical covariance structure for mice in the same tumorgraft line.
RNAseq values were log2-transformed for analysis; Raw values depicted in all
graphs. Bar charts depict individual RNA-seq values, while all boxplots have
median centre values. **, p < 0.01; and ****,
p < 0.0001. See Supplementary Fig. 1 for gel source
data.