Figure 1
MCP-1 and IL-8 induces actin stress fibers and alters contractile properties of human TM cells. Primary cultured TM cells we generated were cultured on 2% gelatin-coated glass cover slips, serum-starved for 24 h and were treated with MCP-1 (100 ng/ml) or IL-8 (100 ng/ml) for 2 h or TGF-β 1 (10 ng/ml) for 24 h before staining for F-actin with TRITC-Phalloidin. Representative confocal images of stained cells show an increase in actin stress fibers (red) with MCP-1 and IL-8 treatment (A). MCP-1 and IL-8 treatment also revealed a significant increase in the levels of phosphorylated MYPT1 (B,C) relative to control (CTL) based on immunoblotting and densitometric analyses. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was immunoblotted as a loading control for the cell lysates. The immunoblot data were normalized to GAPDH. Full-length blots/gels are presented in Supplementary Figure S1. *P < 0.05. N = 3, values are mean ± SEM and one-way ANOVA with Dunnett’s multiple comparison was done. MCP-1 monocyte chemoattractant protein, IL-8 interleukin-8, MYPT1 myosin phosphatase targeting subunit 1, GAPDH glyceraldehyde 3-phosphate dehydrogenase.
