Fig. 5

UC-MSCs combined with decitabine more effectively polarized M1 macrophages towards M2 macrophages in vitro. BMDMs were isolated from C57BL/6 and stimulated by LPS and IFN-γ for 12 h (LPS + IFN-γ group). Then, LPS + IFN-γ-induced BMDMs (M1 macrophages) were cultured with 5 × 10⁴ UC-MSCs in a Transwell system for 24 h (MSC group), with decitabine (10 nmol/L) for 48 h (DAC group), or with decitabine for 48 h after coculture with UC-MSCs for 24 h (M+D group). a Quantitative RT-PCR analysis of inflammation-related gene expression in macrophages from five groups. b Representative macrophage phenotype-related gene expression in macrophages by QT-PCR analysis. c Immunofluorescence and quantification of iNOS-positive and Fizz1-positive cells in five groups. Scale bar = 50 μm. Quantification was determined by evaluating cells from at least 5 sections of each slide, at least 3 slides per group. d Immunoblotting analysis of iNOS and Arg-1 expression in macrophages. Relative protein levels are quantified by the ratio of iNOS or Arg-1 to β-tubulin. The results are presented relative to those of the LPS + IFN-γ group. Images of a–d are representatives of three independent experiments. Values are the mean ± SD (n = 3) of three individual experiments, *P < 0.05; **P < 0.01