Extended Data Fig. 7: Fitness of Pik3caH1047R/wt cells depends on Hif1α.
From: Organismal metabolism regulates the expansion of oncogenic PIK3CA mutant clones in normal esophagus
a-d, Protocol (a): Pik3cawt/wt (wild-type) or Pik3caH1047R/wt mutant cells expressing shRNA against Hif1a (shHif1a) or control shRNA (shNT) were mixed with WT-RYFP and cultured for 28 days in minimal medium. b, Hif1α mRNA levels in shHif1a or shNT cells. Each dot is a culture from a different mouse (n = 4 mice). Red lines, median values. Two tailed paired t-test. c, Representative confocal images showing basal layer of an epithelioid culture generated as in a. n = 3 biological replicates. WT-RYFP cells, yellow, DAPI, blue. Scale bar, 20 μm. d, Proportion of WT-RYFP cells at 28 days versus day 0. Each dot is a culture from a different animal. n = 3-7 cultures per condition. Two-tailed paired t-test. e-f, In vitro cell competition of WT-RYFP mixed with Pik3caH1047R/wt cells or uninduced controls from the same mice. Cultures were then treated with HIF1α inhibitor PX-478 (10 µM) or vehicle for 28 days. (f). Each dot is mean of a culture from a different animal (n = 5-11). Bars are SD. Two tailed unpaired t-test.
