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. 2022 Nov 23;49(1):14. doi: 10.3892/or.2022.8451

Figure 5.

Figure 5.

ATF4 KD affects HIF1α and HIF2α expression in acute and chronic hypoxia. PANC-1 cells were treated with RNAiMAX Lipofectamine reagent and small interfering RNA to transiently KD ATF4, HIF1α or HIF2α. The cells were placed in hypoxia (0.2% oxygen) or normoxia (21% oxygen) for 16 or 48 h. All cells with KD were placed in hypoxia. (A and B) RT-qPCR was conducted to determine HIF1α after (A) 16 and (B) 48 h of hypoxia. (C) HIF2α mRNA levels were measured using RT-qPCR after ATF4 KD in acute or chronic hypoxia. (D and E) Western blot analysis was conducted to determine the protein expression levels of HIF1α after (D) 16 or (E) 48 h of hypoxia. Fold changes are shown relative to normoxic cells. Error bars represent the mean ± SD for n=3. Fold changes were analyzed by one-way ANOVA followed by Bonferroni statistical hypothesis test. Fold changes for RT-qPCR are shown relative to normoxic cells. Error bars represent the mean ± SD for n=9. Fold changes were analyzed by one-way ANOVA followed by Bonferroni statistical hypothesis test. **P<0.01, ***P<0.001 and ****P<0.0001. ATF4, activating transcription factor 4; HIF, hypoxia-inducible factor; KD, knockdown; RT-qPCR, reverse transcription-quantitative PCR.