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. Author manuscript; available in PMC: 2017 Jul 6.
Published in final edited form as: Nature. 2016 Sep 5;539(7627):107–111. doi: 10.1038/nature19795

Figure 2.

Figure 2

On-target effects of PT2399 on transcription and soft agar growth. a, Immunoblots of HIF2α−/− 786-O cells (sgHIF2α #6) infected with a lentivirus encoding V5-tagged HIF2α (wild-type), HIF2α (S304M), or empty vector. b, Levels of the indicated mRNAs, normalized to beta actin mRNA, and then to DMSO treatment, in cells from (a) treated with PT2399 or DMSO for 48 hours; n=3. c, d, Soft agar colonies formed by 786-O cells infected with a lentivirus encoding HIF2α S304M (d) or the empty vector (c) in the presence of PT2399 at the indicated concentrations for 21 days; n=3 biological replicates. e, f, Soft agar colonies formed by 786-O clones as in (Extended Data Figure 2a, b); n=3 biological replicates. In f, the 786-O HIF2α−/− cells (sgHIF2α #6) were superinfected with a lentivirus encoding a sgRNA-resistant HIF2α cDNA (HIF2α) or with the empty vector (EV). g, Quantification of soft agar colony formation in (c–f). Data as mean ± s.e.m. *P<0.05, **P<0.01 and ***P<0.001 by two-tailed Student’s t-tests (b, g).