miR-129-5p Regulates the Immunomodulatory Functions of Adipose-Derived Stem Cells via Targeting Stat1 Signaling

Figure 3

iNOS expression in and inhibition of T cell proliferation by ASCs with miR-129-5p knockdown. Both NC- and miR-129-5p inhibitor-transduced ASCs were treated with TNFα plus IFNγ for 12 h at increasing concentrations. (a) mRNA levels of inflammatory cytokines in NC- and miR-129-5p inhibitor-transduced ASCs were determined by quantitative RT-PCR. (b) After treatment with TNFα plus IFNγ for 24 h, the amount of nitric oxide in the supernatant of NC- and miR-129-5p inhibitor-transduced ASCs was examined by a Griess test. (c) Both NC- and miR-129-5p inhibitor-transduced ASCs were treated with LPS (1.5 μg/mL) for 12 h. mRNA levels of inflammatory cytokines were then determined by quantitative RT-PCR. (d) CD3⁺ T cells isolated from murine spleens were labeled with CFSE and stimulated with PMA (50 ng/mL) plus ionomycin (1 μg/mL) for 24 h and then cocultured with NC- or miR-129-5p inhibitor-transduced ASCs at different ratios (ASC : T cell). After 48 h, cells were analyzed by flow cytometry for T cell proliferation as indicated by reduced CFSE fluorescence intensity. and .

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