Fig. 2

Knockdown of MRF in MSCs inhibits monocyte recruitment and M1 macrophage polarization but facilitates M2 polarization. A The downregulation of MRF expression was verified by qRT–PCR after knockdown (n = 3). B The number of migrated monocytes induced by MSCs was decreased after MRF knockdown (n = 3). C The number of migrated monocytes induced by MSC culture supernatant was decreased after MRF knockdown (n = 3). D The MCP1 mRNA level in MSCs was measured by qRT–PCR after MRF knockdown under the conditions of coculture with or without CD14+ monocytes (n = 3). E The MCP1 protein level in MSCs was detected by western blotting, and the quantitative results were normalized to the result for β-tubulin (n = 3). F MCP1 levels in MSC culture supernatants were quantified via ELISA (n = 3). G The MFI of HLA-DR (left panel) and CD206 (right panel) was detected in CD68-positive cells via flow cytometric analysis (n = 3). H Histogram showing the MFI of HLA-DR among the groups cultured without MSCs, with MSCs and with MRF-knockdown MSCs. I Histogram showing the MFI of CD206 among the groups cultured without MSCs, with MSCs and with MRF-knockdown MSCs. The data are presented as the mean ± SD; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns not significant. All experiments were independently repeated three times