Fig 1: Exosome-Rifampicin activated phosphorylation of Drp1 to induce mitochondrial apoptosis. (A and B) The effects of Exosome-Rifampicin on mitochondrial fusion/fission protein expression in OS cells. Representative immunoblot of the protein levels of p-Drp1, Drp1, Fis1, Opa1, Mfn1 and Mfn2. n = 3. (C) Detection of Mitotracker fluorescence in OS cells by flow cytometry. n = 4. (D) ATP levels in OS cells. n = 6. (E) Immunoblot analysis of Nudfs1, Sdha, Uqcrc1 and ATP5A. n = 3. (F) Apoptosis detection by flow cytometry. n = 4. (G) Fluorescence microscopy of MitoSOX fluorescence in OS cells. n = 4. *p <0.05, **p <0.01, ***p <0.001.
Fig 2: Model of neuritic degeneration induced by polybrene.In response to polybrene, a rapid extracellular Ca2+ influx occurs, which initiates two independent mechanisms involved in neural degeneration: DRP1-dependent mitochondrial fragmentation and ER-mitochondria interface remodeling
Fig 3: Hsp22 regulates PGC1α via AMPK signaling pathway in rats after SAH Beam balance scores, Modified Garcia scores and Brainwater content in various groups. n = 6 per group. (B) Representative photomicrographs of TUNEL staining and quantitative analyses in the indicated groups. n = 4 per group. Scale bar = 100 μm. (C) Typical photomicrographs showing double immunofluorescence staining of PGC1α (green) and NeuN (red) in diverse experimental groups. n = 4 per group. Scale bar = 50 μm. (D) Western blot images and quantitative analyses of p-AMPK/AMPK, PGC1α, Drp1, Nrf1, TFAM, UCP2, Cleaved caspase-3/Caspase-3, Bcl2, Bax, Cytosolic and mitochondrial cytochrome c. n = 6 per group. Bars represent mean ± SD. **P < 0.01, *P < 0.05 vs. Sham group. ##P < 0.01, #P < 0.05 vs. SAH + Vehicle group. &&P < 0.01, &P < 0.05 vs. SAH + hsp22+scramble siRNA. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
Fig 4: The beneficial effects of Tat-Beclin 1 on mitochondrial homeostasis in adult mouse ventricular cardiomyocytes (AMVMs) depend on autophagy. Mouse model of inducible cardiomyocyte-specific ATG7 knockout mice (ATG7 cKO (KO)) were generated. ATG7F/F treated with tamoxifen injection (WT) were used as control. AMVMs were treated with TS or TB for 2 h, DNA and RNA were extracted for qPCR and qRT-PCR, respectively. (A,B), representative Western blot, and quantification of ATG7 expression and LC3-II level in AMVMs. n = 3. WT vs. KO, ATG7, p = 0.0151. LC3-II, p = 0.002. (C), ATG7 gene expression in AMVMs. n = 5–6. WT vs. KO, p = 0.0073. (D), PGC1α gene expression in AMVMs. n = 5–6. TS vs. TB, WT, p = 0.0212. KO, p = NS. (E,F), mitochondrial dynamics fission-related gene expression in AMVMs. n = 5–6. TS vs. TB, Drp1, WT, p = 0.048. KO, p = NS. Fis1, WT, p = 0.0365. KO, p = NS. (G–I), mitochondrial dynamics fusion-related gene expression in AMVMs. n = 5–6. TS vs. TB, Opa1, WT, p = 0.3713. KO, p = NS. Mfn1, WT, p = 0.0104. KO, p = NS. Mfn2, WT, p = 0.0243. KO, p = NS. (J), mtDNA copy number in cardiomyocytes (AMVMs) were analyzed by qPCR of ATP6. n = 5–6. TS vs. TB, WT, p = 0.0494. KO, p = NS. TS, Tat-Scrambled; TB, Tat-Beclin 1; NS (not significant).
Fig 5: The beneficial effects of Tat-Beclin 1 on mitochondria in mouse hearts depend on autophagy. ATG7 cKO (KO) (αMHC-merCremer+; ATG7F/F injected with tamoxifen) were generated. ATG7F/F treated with tamoxifen injection (WT) were used as control. Mice were treated with Tat-Scrambled or Tat-Beclin 1 for 24 h, and the left ventricle was isolated for analysis. (A), ATG7 gene expression in mouse heart. n = 9–10. WT vs. KO, p = 0.0002. (B), PGC1α gene expression in mouse heart. n = 4–6. TS vs. TB, WT, p = 0.023. KO, p = NS (not significant). (C,D), mitochondrial dynamics fission-related gene expression in mouse heart. n = 4–6. TS vs. TB, Drp1, WT, p = 0.0421. KO, p = NS. Fis1, WT, p = 0.0216. KO, p = NS. (E–G), mitochondrial dynamics fusion-related gene expression in mouse heart. n = 4–6. TS vs. TB, Opa1, WT, p = 0.0205. KO, p = NS. Mfn1, WT, p = 0.0121. KO, p = NS. Mfn2, WT, p = 0.0459. KO, p = NS. (H), mtDNA copy numbers in mouse hearts were analyzed by ATP6 qPCR. n = 4–6. TS vs. TB, WT, p = 0.0095. KO, p = NS. TS, Tat-Scrambled; TB, Tat-Beclin 1; NS (not significant).
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