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. 2020 Jan 1;10(1):17–35. doi: 10.7150/thno.36930

Figure 5.

Figure 5

Overexpression of MSR1 in RAW264.7 cells promotes mitochondrial OXPHOS and M2-like polarization in the co-culture system. (A) OCR of blank (BL), vector (Vec), and MSR1-overexpressed (OE) RAW264.7 cells after co-culture were evaluated using a Seahorse Bioscience XFp analyzer. (B) Basal respiration, ATP production, respiratory capacity, and respiratory reserve were determined in BL, Vec, and MSR1 OE RAW264.7 cells with or without co-culture. Values are expressed as mean ± SD, **p < 0.01, ***p < 0.001, ns indicates no significance. (C) IF staining of BL, Vec, and OE RAW264.7 cells after co-culture for the M1-like biomarker (iNOS) and M2-like biomarker (CD206). Bar = 50 μm. (D) mRNA expression levels of M2-like macrophage marker genes (CD206 and CD163) in indicated groups were analyzed by qPCR. Values are expressed as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, ns indicates no significance. (E and G) Flow cytometry analysis of RAW264.7 cells from different groups after co-culture with BMSCs. Dot plots represent F4/80 and iNOS staining of (E) and F4/80 and CD206 staining of RAW264.7 cells (G). (F and H) percentages of F4/80+ iNOS+ (F) and F4/80+ CD206+ (H) in RAW264.7 cells in different groups were calculated. Values are expressed as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, ns indicates no significance.