Fig. 7.

DOT1L regulates the expression and activation of JAK3 in macrophages. a Co-immunofluorescence photomicrographs illustrated co-staining of DOT1L and CD163 in the peritoneum from patient and mouse with peritoneal fibrosis induced by PDF. Arrows indicated CD163-positive cells. b Raw264.7 were transfected with scrambled siRNA and DOT1L siRNA for 6 h, and then incubated with 10ng/ml IL-4 for an additional 36 h before being harvested for analysis. Cell lysates were subjected to immunoblotting analysis with antibodies against TYK2, JAK1, JAK2, JAK3, Src and GAPDH. c, d Raw264.7 were transfected with Flag-DOT1L pcDNA3.0 plasmid or vector for 0, 18, 36 h and then subjected to immunoblotting analysis with antibodies against DOT1L, H3K79me2, Histone H3, TYK2, JAK1, JAK2, JAK3, p-JAK3 and GAPDH. e ChIP-qPCR analysis of H3K79me2 enrichment in the JAK3 promoter region. f 10ng/ml IL-4 treated Raw264.7 were subjected to immunoprecipitation with IgG or DOT1L antibody, followed by JAK3 and DOT1L immunoblotting. Input lysates were analyzed by JAK3, DOT1L and GAPDH immunoblotting. g Immunofluorescence co-staining of DOT1L and JAK3 in IL-4-treated Raw264.7