Fig. 2.
SP upregulates gene expression and protein secretion of various ECM components in corneal fibroblasts during the onset on fibrosis in vitro. a RT-qPCR showed that 10−5M SP upregulates COL1A1 gene expression in corneal fibroblasts both 2 days and 4 days after treatment and induction of fibrosis. Supernatants collected 2 days and 4 days after induction of fibrosis and treatment with 10−5M SP were subjected to pro-collagen I ELISA. Secretion of pro-collagen I was increased in supernatants collected from SP-treated cells both 2 days and 4 days after treatment. b 10−5M SP upregulated COL3A1 gene expression in corneal fibroblasts both 2 days and 4 days after treatment and induction of fibrosis. Supernatants collected 2 days and 4 days after induction of fibrosis and treatment with 10−5M SP were subjected to collagen III ELISA. SP treatment had no effect on collagen III secretion at either 2 days or 4 days after treatment. c 10−5M SP upregulated COL5A1 gene expression in corneal fibroblasts both 2 days and 4 days after treatment and induction of fibrosis. Supernatants collected 2 days and 4 days after induction of fibrosis and treatment with 10−5M SP were subjected to collagen V ELISA. SP treatment had no effect on collagen V secretion at either 2 days or 4 days after treatment. d 10−5M SP downregulated LUM gene expression in corneal fibroblasts at day 2, but 4 days after treatment and induction of fibrosis the LUM gene expression was increased. Supernatants collected 2 days and 4 days after induction of fibrosis and treatment with 10−5M SP were subjected to lumican ELISA. SP treatment had no effect on lumican secretion at day 2; however, it increased lumican secretion 4 days after treatment. Values are means ± SD. n.s. (not significant); *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001