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. 2021 Sep;9(18):1435. doi: 10.21037/atm-21-4080

Figure 4.

Figure 4

The protective effects of AS-IV on hypoxic injury were suppressed by HIF-1α downregulation. (A,B) RT-qPCR and western blot analysis of HIF-1α expression in H9c2 cells transfected with si-HIF-1α or control si-NC. (C,D) HIF-1α protein levels after siRNA transfection, and the effects of hypoxia and AS-IV treatment were determined by immunofluorescence and western blot analyses. (E) Cell survival was measured using the CCK-8 assay. (F) Cell death was assayed by calcein-AM/PI staining. (G) Apoptosis was assayed by flow cytometry. (H) Western blot analysis of the levels of the apoptotic proteins cleaved caspase-3, Bcl-2, and Bax. β-actin was used as the loading control. Results are mean ± SD of three independent experiments performed in duplicate. *P<0.05 or **P<0.01 vs. corresponding group. AS-IV, astragaloside IV; HIF-1α, hypoxia-inducible factor 1α; RT-qPCR, quantitative reverse transcription PCR; si-HIF-1α, HIF-1α siRNA; si-NC, negative control siRNA; CCK-8, Cell Counting Kit-8; PI, propidium iodide.