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. 2020 Jun 5;10(16):7319–7334. doi: 10.7150/thno.44054

Figure 5.

Figure 5

SFN dependeds on the Nrf2-miR-93-TLR4/IRF1 axis to suppress TLR4 and IRF1 expression and promote M2Mϕ polarization in vitro. Western blot (A) and qPCR (B) analyses of Nrf2, TLR4, IRF1, iNOS, and ARG-1 levels in BMDMs treated with SFN and/or miR-93 inhibitor. GAPDH was used as an internal control. (C) The distribution of iNOS (green) and ARG-1 (red) in BMDMs treated with SFN and/or miR-93 inhibitor according to immunofluorescence (200× magnification; scale bar: 20 µm). (D) Fluorescence microscopy analysis of BMDM phagocytic ability. COM crystals were labeled with an Alexa Fluor 488-conjugated IgG and directly cultured with treated BMDMs (200× magnification; scale bar: 20 µm). Flow cytometric analysis of BMDM polarization (E) and TECs necrosis (F) in co-cultured cells treated with SFN and/or miR-93 inhibitor. Data represent the mean ± standard error (SE) of three independent experiments. *P < 0.05; **P < 0.01, as determined by one-way ANOVA (B-G).