Figure 7.

ATF4, HIF1α and HIF2α KD decrease cell viability in chronic hypoxia. (A and B) PANC-1 cells were transiently transfected and seeded into 96-well plates and incubated in hypoxia or normoxia for (A) 16 or (B) 48 h. Cell viability was measured by CellTiter Glo luminescence. The negative control were PANC-1 cells exposed to 0.2% oxygen with transfection reagent but no KD small interfering RNA. Error bars represent the mean ± SD for n=5. The relative luminescence unit (RLU) were analyzed by one-way ANOVA followed by Bonferroni statistical hypothesis test. (C) Colony formation assays were performed by transiently transfecting PANC-1 cells and seeded into plates. The plates were placed in either normoxia or hypoxia for 16 or 48 h. The cells were fixed, stained and colonies were counted. Error bars represent the mean ± SD for n=3. **P<0.01, ***P<0.001 and ****P<0.0001. ATF4, activating transcription factor 4; HIF, hypoxia-inducible factor; KD, knockdown.