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. 2023 Feb 8;14(1):2171636. doi: 10.1080/21505594.2023.2171636

Figure 2.

Figure 2.

Resistin inhibits claudin-5 and occludin expression in PAECs. (a, b, c) PAECs were co-cultured with wild-type or resistin knockout PAMs, and the PAMs were infected by H. parasuis (100 MOI) for 2 h. mRNA levels of claudin-5, occludin, JAM-1, JAM-2, and VE-cadherin in PAECs were determined using qRT-PCR (a, b), and relative protein levels were detected using western blot analysis (c). (d, e, f) PAECs were stimulated or unstimulated with recombined resistin protein (2.5, 5, or 0 nM) for 2 h. Cells were lysed, mRNA levels of claudin-5 and occludin were analysed using qRT-PCR (d, e), and protein levels were analysed using western blot analysis (f). The mRNA level of each gene was standardized to GAPDH. The antibodies against claudin-5, occludin, JAM-1, JAM-2, and VE-cadherin were diluted at 1/1000, and the antibody against β-actin was diluted at 1/50000. β-actin served as a loading control, and the relative protein level was calculated with ImageJ software and normalized to β-actin. **p<0.01 compared with the wild-type group (a, b, c) or the untreated group (d, e, f). ##p<0.01 compared with the untreated group. Error bars represent the mean ± SEM (n = 3).