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. 2023 Feb 28;14:1124269. doi: 10.3389/fimmu.2023.1124269

Figure 2.

Figure 2

Increased MHC class II expression by ET-1 incubation in vitro. (A) EA.hy926 endothelial cells were incubated with 1, 10, and 100 nM ET-1. (B) EA.hy926 cells were incubated with 100nM ET-1 (dark grey bar) or preincubated with 10µM BQ788, a selective blocker of ET-1 type B receptors (white bar) for 24 h at 37°C and compared to vehicle treatment (light grey bar). Gene expression was quantified by qRT-PCR using TaqMan probes. (C) CIITA Chromatin Immunoprecipitation. CIITA is associated with MHC class II promoter in EA.hy926 cells treated with 100 nM ET-1 for 24 hours at 37°C. Lysates were immunoprecipitated with antibodies against CIITA, and DNA was isolated and analyzed by real-time PCR. Values were normalized to the total amount of promoter DNA added to the reaction (input). Input = 5% of total cell lysate. Data represent mean ± SD (n = 3). *p < 0.05, **p < 0.01, 100nM ET-1 versus 100nM ET-1 with 10µM BQ-788..