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Fig. 6 | Molecular Medicine

Fig. 6

From: Phosphorylation of BCL2 at the Ser70 site mediates RANKL-induced osteoclast precursor autophagy and osteoclastogenesis

Fig. 6

BCL2 mutation at S70 enhanced the coimmunoprecipitation of BCL2 and Beclin1 in OCPs. AC The transfected OCPs were treated with RANKL for 2 h in α-MEM with 1% FBS. Cell lysates were extracted for Co-IP with anti-BCL2, anti-Beclin1 or anti-BAX antibody, and subsequently, precipitates were observed using western blotting assays with anti-Beclin1, anti-BAX or anti-BCL2 antibody, respectively. D The transfected OCPs were treated with RANKL for 24 h in α-MEM with 2.5% FBS. The mitochondrial membrane potential was detected by measuring the ratio of red/green fluorescence intensity using JC-10 kit, and quantitatively analysed using flow cytometry. E The percentages of cells in each quadrant are shown in the histograms according to the results in D. The larger proportion of cells in H2 quadrant indicates that mitochondrial membrane potential is higher; The larger proportion of cells in H4 quadrant indicates that mitochondrial membrane potential is lower. The experiments were replicated at least three times. Data are presented as the mean ± SEM from three independent experiments. ***P < 0.001. IP the antibody for immunoprecipitation, IB the antibody for immunoblot

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