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Fig. 7 | Stem Cell Research & Therapy

Fig. 7

From: SPION-MSCs enhance therapeutic efficacy in sepsis by regulating MSC-expressed TRAF1-dependent macrophage polarization

Fig. 7

Knockout of TRAF1 weakens the immunomodulatory effect of SPION-labelled/pretreated MSCs on macrophages. A, B Immunoblotting analysis of TRAF1, p65, p-p65, cox2, IL-10 and GAPDH in the RAW 264.7 cells transfected with si-TRAF1 for 24 h and then stimulated with SPIONs for 24 h. C, D Representative flow cytometry plots showing the gating strategy used to determine total macrophages, M1 macrophages (iNOS + CD206 −) and M2 macrophages (iNOS-CD206 +). E Mid-grade sepsis was induced by CLP in male C57BL/6 mice. Sham-operated mice underwent the same procedure without ligation and puncture of the caecum. PBS or a suspension of SPION-MSCs or siTRAF1-SPION-MSCs (106 cells in 150 μl of PBS). F, G Flow cytometry plots and analysis of the percentage of CD11b + F4/80 + macrophages and the M2 marker CD206 and M1 marker iNOS. H Representative images of western blots to assess the levels of iNOS, IL-10 and Arg1 in the mouse livers. Data with error bars are presented as the mean ± SD. Each panel is a representative experiment of at least three independent biological replicates. Scale bars, 50 mm. *p < 0.05, **p < 0.01, ***p < 0.001 as determined by unpaired Student’s t-test

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