Fig. 4.

AST-pretreated MSCs promote a change in the phenotype of macrophages to M2 by increasing the secretion of PGE2. (A–E) CD163 and CD206 protein expression in RAW264.7 cells after co-cultured MSCs or astilbin (AST) pretreated MSCs. (A)Western blot analysis showing the expression of CD163, CD206 and F4/80. Phenotypic changes of RAW264.7 cells were evaluated by CD163 (B and C) and CD206 (D and E) protein levels. CD163 and CD206 protein levels were normalized to the GAPDH and F4/80 levels. The levels of PTGES mRNA (F) and PGE2 (G) in supernatants were measured using real-time PCR and ELISA, respectively. (H–J) Astilbin-treated MSCs transfected with shRNA-NC increased CD163 protein levels, whereas astilbin-treated MSCs transfected with shRNA-PTGS2 reversed this upregulation. (n = 6 in each group *p < 0.05; ***p < 0.001) Corresponding uncropped full-length gels and blots are presented in Additional file 1