Figure 1

IL-17A derived from SSc patient serum promotes the proliferation, collagen synthesis and secretion of SSc patient-derived DVSMCs. (A) DVSMCs were treated with different doses of IL-17A for 24 h, 48 h, 72 h, and cell proliferation was tested using cell counting kit-8. (B) The cells were treated with the serum of SSc patients for 24 h, 48 h, 72 h, and cell proliferation was tested using CCK8. (C) The activity of collagen1α1 or collagen3α1 proximal promoter was test using a dual luciferase reporter gene assay after being treated with different doses of IL-17A for 24 h. (D) The activity of collagen1α1 or collagen3α1 proximal promoter was also detected after being treated with the serum of SSc patients and healthy subject for 24 h. Data were represented as mean ratios of Firefly to Renilla luciferase activity. (E) The cells were cultured in the indicated doses of IL-17A for 24 h, the gene expression of collagen 1 and collagen 3 was measured using real-time RT-PCR analysis. (F) The cells were stimulated with the serum of SSc patients and healthy subjects for 24 h, and the gene expression of collagen 1 and collagen 3 was measured using real-time RT-PCR analysis. (G) The cells were treated with different doses of IL-17A for 24 h, the concentration of collagen 1, collagen 3 was detected using ELISA. (H) The cells were treated with the serum of SSc patients and healthy subjects for 24 h, and the concentration of collagen 1, collagen 3 was also detected by ELISA. (I, J) The cells were treated with different doses of IL-17A, serum of SSc patients or healthy individuals for 24 h, and the expressions of collagen 1, collagen 3, IL-17RA and α-SMA were measured using Western blot. GAPDH was used as a loading control. The experiment was repeated three times, and the data are presented as means ± standard deviation.