FIS1 and MiD49 as the adaptors of DRP1 mediates PRRSV-induced mitochondrial fission.
<p>(A-B) Marc-145 cells were transfected with control siRNA or siRNA targeting FIS1 (A) or MiD49 (B) for 24 h, and then were either mock-infected or infected with PRRSV (0.1 MOI) for an additional 24 h. The cells were stained with MitoTracker Red, PRRSV-N antibody, and DAPI, followed by confoc...
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2025
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| Summary: | <p>(A-B) Marc-145 cells were transfected with control siRNA or siRNA targeting FIS1 (A) or MiD49 (B) for 24 h, and then were either mock-infected or infected with PRRSV (0.1 MOI) for an additional 24 h. The cells were stained with MitoTracker Red, PRRSV-N antibody, and DAPI, followed by confocal microscopy. Quantitative analysis of mitochondrial length is presented for 25 cells per group (right panel). (C-D) Marc-145 cells were mock-infected or infected with PRRSV (MOI = 0.1) for 24 h. The cells were lysed and precipitated using anti-FIS1 or anti-MiD49 antibodies. The whole-cell lysates (WCL) and immunoprecipitated proteins were analyzed using specific antibodies. Anti-Rabbit immunoglobulin G (IgG) antibody was used as a negative control. (E and F) After 24 hours of siRNA FIS1 (E) and MiD49 (F) transfection, the cells were either mock-infected or infected with PRRSV (0.1 MOI) for an additional 24 hours, and then harvested for cytoplasmic and mitochondrial fractionation. Then samples were subjected to western blot analysis. The levels of proteins were normalized to Tom20 or α-tubulin. Data are expressed as means ± SD, n = 25 in A and B. *p<0.05; **p < 0.01; ***p < 0.001. The data are representative of results from three independent experiments.</p> |
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