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. 2021 Mar 16;48:101214. doi: 10.1016/j.molmet.2021.101214

Figure 4.

Figure 4

RYGB surgery modulated microglia and POMC neuron interaction by a humoral factor. (A and B) BV2 cells were 4-h treated with plasma (2%) collected from RYGB- and sham-operated DIO and BWM rats at 12 weeks post-operatively. (A and B) NF-kB nuclear translocation (upper panel) in cells was assessed by immunofluorescent staining with an anti–NF–kB antibody (red) and DAPI (blue), while morphology was visualized by IBA1 fluorescent staining (lower panel), scale bar, 50 μm (A) and quantification by cell counting of immunofluorescent staining (B). (C) BV2 cells were pretreated by 100 nM TAK242 (+) or vehicle (−) for 1 h before further incubation with medium containing 2% rat plasma for 4 h. The mRNA expression of Il-1b, Il18, Tnfa Tlr4, Myd88, and Cd14 was analyzed by qPCR. (D) Murine hypoPOMC neurons were incubated for 4 h with conditioned medium (CM) obtained from BV2 cell culture that was exposed to rat plasma (2%, 4 h). The mRNA expression of Pomc and Obrb was assessed by qPCR. (E) POMC neurons were incubated with the same CM for 24 h and cell viability was measured by a CyQUANT XTT Cell Viability Assay. Data are presented as mean ± SEM (n = 3–4) with individual datapoints. ∗P < 0.05, ∗∗P < 0.01, and ∗P < 0.001 for the effect among non-TAK242 pretreated cultures; ###P < 0.001 for the effect of TAK242 vs vehicle pretreated cultures for C; ∗∗P < 0.01 and ∗∗∗P < 0.001 for the effect of any indicated comparison for D and E.