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. 2022 Jun 22;5(11):e202101264. doi: 10.26508/lsa.202101264

Figure 6. Hypoxia inducible factor-1⍺ (HIF-1⍺) negatively regulates AREG expression in pulmonary arterial endothelial cells (PAECs) under hypoxic conditions.

Figure 6.

(A) Heat map showing BRCA1, HLX, NCOA6, PHB2, RRP1B, TAF4, TP63, and VAV2 expression assessed by qRT-PCR in normoxic and hypoxic PAECs. (B) The binding of HIF-1⍺ to the NCOA6, PHB2, and RRP1B promoter regions in PAECs was assessed by ChIP qPCR. (C, D, E, F, G) PAECs were transfected with scrambled siRNA (siCTL) or siRNA against NCOA6 (siNCOA6), PHB2 (siPHB2), or RRP1B (siRRP1B) and placed in hypoxia for 24 h. (C) Apoptotic PAECs were quantified by measuring caspase 3+ cells and caspase 3 MFI by flow cytometry. (D) PAECs were plated on Matrigel, and tube formation was assessed. (E, F) AREG (E), IFNB, IL1B, IL6, and TNFA (F) expression was assessed by qRT-PCR. (G) PAECs were transfected with either scrambled siRNA (siCTL) or siRNA against HIF1A (siHIF1A) and placed in hypoxia or normoxia for 24 h. NCOA6, PHB2, and RRP1B expression was quantified by qRT-PCR. n = 5 replicates per condition. Data are shown as mean. *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001.