Fig. 2

HIF-1α induces CD146 expression in PASMCs. a, b Relative mRNA levels of CD146 in hPASMCs exposed to normoxia or hypoxia for indicated times. c CD146 protein expression in hPASMCs exposed to 21% or 1% O₂ for indicated times. d, e 293 T cells were transfected with siRNA targeting HIF1A, HIF2A, or TP53 for 24 h before exposed to 1% or 21% O₂ for another 24 h. CD146 expression at mRNA d or protein e level was measured. f, g Human PASMCs were treated with hypoxia, CoCl₂ (100 μM), DMOG (100 μM) or DFX (50 μM) for 24 h before CD146 mRNA f or protein g level was measured. h–k Human PASMCs were transfected with HIF-1α-expressing plasmid h, i or HIF-1α-siRNA j, k for 24 h before CD146 expression was measured. l Design of luciferase reporter vector containing human CD146 promoter that includes an HRE or a mutant HRE (mHRE). m Dual luciferase assay of putative HIF-1α-binding sites in CD146 promoter. The luciferase activity in pGL3 transfected construct was set as 1. n ChIP using ChIP-grade HIF-1α antibody and quantification of the enrichment of HIF-1α binding to CD146 promoter after hypoxia stimulation (n = 3 per group). o, p Luciferase assay of HRE-WT reporter in human PASMCs transfected with HIF-1α siRNAs o, or treated with HIF-1α inhibitor digoxin or acriflavine. p Reporter activity was measured and plotted after normalizing with respect to Renilla luciferase activity. n = 3 n, n = 4 a, b, d, f, h, j, or n = 5 m, o, p biological replicates for each group. All WB represent data from three c, e, g, i, k independent experiments. In all statistical plots, the results are expressed as mean ± s.e.m. *P < 0.05, *P < 0.01, ***P < 0.001. Two-tailed Student’s t test. Source data are provided as a Source Data file