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. 2020 Sep 29;124(2):425–436. doi: 10.1038/s41416-020-01067-1

Fig. 5. G3BP1 interacts with YWHAZ to sequester the apoptosis regulator Bax in the cytoplasm.

Fig. 5

a Representative images of the co-localisation of G3BP1 and YWHAZ in MGC80-3 cells. Cells were incubated with anti-G3BP1 (1:50) and anti-YWHAZ (1:50) antibodies. Scale bar: 25 µm. b Co-immunoprecipitation (co-IP) experiments in MGC80-3 cells were performed using anti-G3BP1 or anti-YWHAZ antibodies. IgG was used as a negative control. c IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with Flag-YWHAZ together with the indicated constructs of HA-G3BP1. d Co-IP experiments in MGC80-3 cells transfected with scramble or siG3BP1 were performed using anti-YWHAZ antibody or anti-rabbit IgG as control. e Left: IB analysis of cell cytoplasm and mitochondria fractions separated from MGC80-3 cells transfected with scramble or siG3BP1. Right: The quantification of relative Bax protein expression. Data were shown as mean ± SD of three independent experiments. ***P < 0.001. f Representative images of Bax localisation in MGC80-3 cells transfected with scramble or siG3BP1 under the treatment of capecitabine (64 μg/mL) for 24 h. Cells were fixed and stained by using anti-Bax (1:50) or anti-COX IV (1:100) antibodies. COX IV staining was indicated as the localisation of mitochondria. Scale bar: 25 µm. g The statistical results of colony formation assay of MGC80-3 cells transfected with the indicated siRNAs under capecitabine (16 μg/mL, 2 weeks) or oxaliplatin (2 μg/mL, 2 weeks) treatment. Data were shown as mean ± SD of three independent experiments. ***P < 0.001. h The proposed model for G3BP1 in regulating tumour apoptosis and chemoresistance in gastric cancer. Scr scramble, IP immunoprecipitation, IB immunoblotting, WCL whole-cell lysate, cyto cytoplasm, mito mitochondria, SGs stress granules.