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Fig. 5 | Journal of Biomedical Science

Fig. 5

From: N-terminal functional domain of Gasdermin A3 regulates mitochondrial homeostasis via mitochondrial targeting

Fig. 5

C-Gsdma3 blocks the cytotoxicity induced by Gsdma3 mutants. (a) Immunoblot analysis of HEK293T cells transfected with the N-Gsdma3/flag, Rim3/flag, M1Btlr/V5, or Gsdma3-I359N/V5 plasmid together with an increasing amount of the C-Gsdma3/mh plasmid. The anti-Gsdma3 antibody is more sensitive than the anti-myc-tag antibody. β-actin was used as a loading control. The asterisk indicates a non-specific band due to the long exposure. (b) Quantification of the immunoblots from (a). The bar graph shows the normalized densitometry levels relative to the sample co-transfected with 2 μg of the C-Gsdma3/mh plasmid. (c, d) Flow cytometry analysis of the 7AAD and DiIC1(5) staining in HEK293T cells transfected with different combinations of plasmid constructs as indicated. The bar graphs represent fractions of dead (7AAD+) cells and mitochondrial membrane potential depolarization (DiIC1(5)low) cells (mean + s.d., n = 3 in duplicates) relative to those in N-Gsdma3- or Rim3 + Gsdma3-trasnfected cells. pIRES vector-transfected cells were used as the staining baseline.. *, P < 0.05; **, P < 0.01; ***, P < 0.001

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