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. 2022 Jun 21;10:925971. doi: 10.3389/fcell.2022.925971

FIGURE 3.

FIGURE 3

Cx43 knockdown reduces inflammatory responses of macrophages co-cultured with dying adipocytes (A) Live imaging of RAW264.7 cells transfected with Gja1 siRNA or controls and co-cultured with dying adipocytes (dACs) for 12 h. Adipocytes were tagged with C12-BODIPY (red) and macrophages were stained with DiO (Green) (n = 3) (B) qPCR analysis of Gja1 and inflammatory cytokine markers in RAW264.7 cells transfected with Gja1 siRNA or controls and co-cultured with dACs for 24 h (n = 6) (C) Extracellular ATP level in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages treated with Gja1 siRNA or control for 30 min (n = 3) (D) qPCR analysis of inflammatory cytokines in RAW264.7 cells transfected with Gja1 siRNA or controls, and co-cultured with dACs in the presence or absence of apyrase (2U/ml) (n = 3) (E) Schematic diagram illustrating the experimental method used for F. Conditioned media obtained from control RAW264.7 cells co-cultured with dACs were transferred into Gja1KD RAW264.7 cells (F) qPCR analysis of inflammatory cytokine markers in Gja1 KD RAW264.7 cells co-cultured with dACs (GjaKD + dAC) or exposed to the conditioned media obtained from the control co-cultures (Gja1KD + CM) (n = 3). Unpaired, two-tailed t-tests (*p < 0.1, **p < 0.01, ***p < 0.001). Each point represents biological replicate. Data are presented as mean ± S.E.M.