Fig. 6

Effects of CCL8 on collagen production of fibroblasts in vitro. A Immunofluorescence staining of CCR8 in a mouse fibroblast cell line (NIH/3 T3). Upper panels: Alexa Fluor 546-conjugated anti-mouse CCR8 antibody staining. Lower panels: isotype control antibody staining. Original magnification × 40 (scale bar, 50 μm). DAPI, 4′6-diamino-2-phenylindole. B mRNA expression levels of Col1a2 in fibroblasts treated with recombinant mouse CCL8 for 24 h. C Total collagen protein production from fibroblasts treated with recombinant mouse CCL8 for 24 h. D Flow cytometric analysis of phosphorylated ERK1/2 (pERK) in fibroblasts treated with recombinant mouse CCL8 at 300 ng/mL of concentration for the indicated times (min). The panels show a representative histogram of each of the 3 wells for the indicated times. E The bar graph shows the rate of pERK1/2-positive cells. F The bar graph shows the mean fluorescence intensity (MFI) of pERK1/2 in fibroblasts treated with recombinant mouse CCL8. The data are presented as means ± SEMs of 5 (B and C) and 3 (E and F) wells in each condition. *p < 0.05, Kruskal-Wallis test