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ROS-Drp1-mitophagy feedback loop controls myogenic differentiation by actin skeleton remodeling <t>mediated</t> <t>MRTF-A/SRF</t> activity. (A-B) Western blot analysis for F-actin, cofilin and p-cofilin was performed at 1 d of differentiation after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5 μM Antimycin A (AA). Mouse Gapdh was used to normalize protein expression. n = 3. (C-D) Western blot analysis for nuclear MRTF-A (n-MRTF-A) expression was performed at 1 d of differentiation after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5 μM AA. Histone H3 was used to normalize protein expression. n = 3. (E-F) Co-immunoprecipitation analysis was performed at 1 d of differentiation to determine the combination of MRTF-A with SRF after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5 μM AA. n = 3. (G-H) Western blot analysis for MyHC was performed at 3 d of differentiation after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5μM AA. Mouse Gapdh was used to normalize protein expression; n = 3. * P < 0.05, ** P < 0.01.
Srf, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ROS-Drp1-mitophagy feedback loop controls myogenic differentiation by actin skeleton remodeling mediated MRTF-A/SRF activity. (A-B) Western blot analysis for F-actin, cofilin and p-cofilin was performed at 1 d of differentiation after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5 μM Antimycin A (AA). Mouse Gapdh was used to normalize protein expression. n = 3. (C-D) Western blot analysis for nuclear MRTF-A (n-MRTF-A) expression was performed at 1 d of differentiation after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5 μM AA. Histone H3 was used to normalize protein expression. n = 3. (E-F) Co-immunoprecipitation analysis was performed at 1 d of differentiation to determine the combination of MRTF-A with SRF after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5 μM AA. n = 3. (G-H) Western blot analysis for MyHC was performed at 3 d of differentiation after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5μM AA. Mouse Gapdh was used to normalize protein expression; n = 3. * P < 0.05, ** P < 0.01.

Journal: Redox Report : Communications in Free Radical Research

Article Title: ROS-Drp1-mitophagy feedback loop regulates myogenic differentiation via actin cytoskeleton remodeling-mediated MRTF-A/SRF axis

doi: 10.1080/13510002.2025.2536400

Figure Lengend Snippet: ROS-Drp1-mitophagy feedback loop controls myogenic differentiation by actin skeleton remodeling mediated MRTF-A/SRF activity. (A-B) Western blot analysis for F-actin, cofilin and p-cofilin was performed at 1 d of differentiation after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5 μM Antimycin A (AA). Mouse Gapdh was used to normalize protein expression. n = 3. (C-D) Western blot analysis for nuclear MRTF-A (n-MRTF-A) expression was performed at 1 d of differentiation after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5 μM AA. Histone H3 was used to normalize protein expression. n = 3. (E-F) Co-immunoprecipitation analysis was performed at 1 d of differentiation to determine the combination of MRTF-A with SRF after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5 μM AA. n = 3. (G-H) Western blot analysis for MyHC was performed at 3 d of differentiation after C2C12 cells were treated with the combination of siDrp1, OE-Pink1 and 5μM AA. Mouse Gapdh was used to normalize protein expression; n = 3. * P < 0.05, ** P < 0.01.

Article Snippet: Antibodies against Tom70 (14528-1-AP), SRF (66742-1-Ig), and MRTF-A for WB/IF were purchased from the Proteintech Group (Wuhan, China).

Techniques: Activity Assay, Western Blot, Expressing, Immunoprecipitation

AAV9-shDrp1 injection blocks skeletal muscle regeneration by regulating actin/MRTF-A/SRF axis. (A) The Drp1 protein expression was detected at 4 w after AAV9-shNC or AAV9-shDrp1 injection. Mouse Gapdh was used to normalize protein expression. n = 3. (B) The weight of TA muscle at 11 d after CTX injection was examined in the AAV9-shNC and AAV9-shDrp1 group. n = 3. (C) H&E staining of TA muscle at 11 d after CTX injection in the AAV9-shNC and AAV9-shDrp1 group. Red arrows indicate the cell nuclei located in the center of muscle fibers. Scale bar, 100 μm. (D) IF staining of dystrophin at 11 d after CTX injection in AAV9-shNC and AAV9-shDrp1 group. White arrows indicate the cell nuclei located in the center of muscle fibers. Scale bar, 100 μm. (E-F) Western blot analysis for MyHC at 3, 5, 7, 9 and 11 d after CTX injection in the AAV9-shNC and AAV9-shDrp1 group. Mouse Gapdh was used to normalize protein expression. n = 3 at each time point. (G-I) Western blot analysis for F-actin, p-cofilin and cofilin was performed at 3 and 5 d after CTX injection in the AAV9-shNC and AAV9-shDrp1 group. Mouse Gapdh was used to normalize protein expression; n = 3. (J-K) Co-immunoprecipitation of MRTF-A and SRF in TA muscle at 5 d after CTX injection in the AAV9-shNC and AAV9-shDrp1 group. n = 3. * P < 0.05, ** P < 0.01.

Journal: Redox Report : Communications in Free Radical Research

Article Title: ROS-Drp1-mitophagy feedback loop regulates myogenic differentiation via actin cytoskeleton remodeling-mediated MRTF-A/SRF axis

doi: 10.1080/13510002.2025.2536400

Figure Lengend Snippet: AAV9-shDrp1 injection blocks skeletal muscle regeneration by regulating actin/MRTF-A/SRF axis. (A) The Drp1 protein expression was detected at 4 w after AAV9-shNC or AAV9-shDrp1 injection. Mouse Gapdh was used to normalize protein expression. n = 3. (B) The weight of TA muscle at 11 d after CTX injection was examined in the AAV9-shNC and AAV9-shDrp1 group. n = 3. (C) H&E staining of TA muscle at 11 d after CTX injection in the AAV9-shNC and AAV9-shDrp1 group. Red arrows indicate the cell nuclei located in the center of muscle fibers. Scale bar, 100 μm. (D) IF staining of dystrophin at 11 d after CTX injection in AAV9-shNC and AAV9-shDrp1 group. White arrows indicate the cell nuclei located in the center of muscle fibers. Scale bar, 100 μm. (E-F) Western blot analysis for MyHC at 3, 5, 7, 9 and 11 d after CTX injection in the AAV9-shNC and AAV9-shDrp1 group. Mouse Gapdh was used to normalize protein expression. n = 3 at each time point. (G-I) Western blot analysis for F-actin, p-cofilin and cofilin was performed at 3 and 5 d after CTX injection in the AAV9-shNC and AAV9-shDrp1 group. Mouse Gapdh was used to normalize protein expression; n = 3. (J-K) Co-immunoprecipitation of MRTF-A and SRF in TA muscle at 5 d after CTX injection in the AAV9-shNC and AAV9-shDrp1 group. n = 3. * P < 0.05, ** P < 0.01.

Article Snippet: Antibodies against Tom70 (14528-1-AP), SRF (66742-1-Ig), and MRTF-A for WB/IF were purchased from the Proteintech Group (Wuhan, China).

Techniques: Injection, Expressing, Staining, Western Blot, Immunoprecipitation

Schematic model of ROS-Drp1-mitophagy feedback loop promotes myogenesis by actin cytoskeleton remodeling-mediated MRTF-A/SRF axis. At the beginning of myogenic differentiation, mitochondrial damage and reactive oxygen species (ROS) levels were elevated. Elevated ROS levels promote Drp1 expression to activate mitophagy and thereby form a ROS-Drp1-mitophagy feedback loop for balancing ROS levels. The ROS-Drp1-mitophagy axis regulates actin skeleton remodeling by promoting p-cofilin expression, allowing MRTF-A to translocate into nuclei and combine with SRF, thereby promoting skeletal muscle differentiation.

Journal: Redox Report : Communications in Free Radical Research

Article Title: ROS-Drp1-mitophagy feedback loop regulates myogenic differentiation via actin cytoskeleton remodeling-mediated MRTF-A/SRF axis

doi: 10.1080/13510002.2025.2536400

Figure Lengend Snippet: Schematic model of ROS-Drp1-mitophagy feedback loop promotes myogenesis by actin cytoskeleton remodeling-mediated MRTF-A/SRF axis. At the beginning of myogenic differentiation, mitochondrial damage and reactive oxygen species (ROS) levels were elevated. Elevated ROS levels promote Drp1 expression to activate mitophagy and thereby form a ROS-Drp1-mitophagy feedback loop for balancing ROS levels. The ROS-Drp1-mitophagy axis regulates actin skeleton remodeling by promoting p-cofilin expression, allowing MRTF-A to translocate into nuclei and combine with SRF, thereby promoting skeletal muscle differentiation.

Article Snippet: Antibodies against Tom70 (14528-1-AP), SRF (66742-1-Ig), and MRTF-A for WB/IF were purchased from the Proteintech Group (Wuhan, China).

Techniques: Expressing