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2-methoxystypandrone (2-MS) inhibits mitochondrial ROS production in senescent fibroblasts. ( A ) Senescent fibroblasts treated with DMSO (0.01%), emodin-1-O-β-glucoside <t>(8</t> <t>μM),</t> <t>emodin-8-glucoside</t> (8 μM), 2-methoxystypandrone (2-MS) (8 μM), or polydatin (8 μM) for 12 days at 4-day intervals. Mitochondrial ROS were measured after staining cells with 30 µM dihydrorhodamine 123 (DHR123. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s post–hoc test, with results considered not significant (n.s.) or significant at ** p < 0.01. Data represent the mean ± S.D., n = 3. ( B ) Senescent fibroblasts treated with DMSO (0.01%) or 2-MS (4, 8, and 12 μM) for 12 days at 4-day intervals. Mitochondrial ROS were measured after staining cells with 30 µM DHR123. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s post–hoc test, with results considered not significant (n.s.) or significant at ** p < 0.01. Data represent the mean ± S.D., n = 3. ( C ) Senescent fibroblasts treated with DMSO (0.01%) or 2-MS (4, 8, and 12 μM) for 12 days at 4-day intervals. DNA tail length was measured using the neutral comet assay. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s post–hoc test, with results considered significant at ** p < 0.01. For each condition, the experiment was repeated three times, yielding three independent sets of data (biological triplicates). Sixteen different samples were analyzed to generate each biological triplicate. Data represent the mean ± S.D.
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2-methoxystypandrone (2-MS) inhibits mitochondrial ROS production in senescent fibroblasts. ( A ) Senescent fibroblasts treated with DMSO (0.01%), emodin-1-O-β-glucoside (8 μM), emodin-8-glucoside (8 μM), 2-methoxystypandrone (2-MS) (8 μM), or polydatin (8 μM) for 12 days at 4-day intervals. Mitochondrial ROS were measured after staining cells with 30 µM dihydrorhodamine 123 (DHR123. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s post–hoc test, with results considered not significant (n.s.) or significant at ** p < 0.01. Data represent the mean ± S.D., n = 3. ( B ) Senescent fibroblasts treated with DMSO (0.01%) or 2-MS (4, 8, and 12 μM) for 12 days at 4-day intervals. Mitochondrial ROS were measured after staining cells with 30 µM DHR123. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s post–hoc test, with results considered not significant (n.s.) or significant at ** p < 0.01. Data represent the mean ± S.D., n = 3. ( C ) Senescent fibroblasts treated with DMSO (0.01%) or 2-MS (4, 8, and 12 μM) for 12 days at 4-day intervals. DNA tail length was measured using the neutral comet assay. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s post–hoc test, with results considered significant at ** p < 0.01. For each condition, the experiment was repeated three times, yielding three independent sets of data (biological triplicates). Sixteen different samples were analyzed to generate each biological triplicate. Data represent the mean ± S.D.

Journal: Antioxidants

Article Title: 2-Methoxystypandrone from Polygonum cuspidatum Rejuvenates Senescence by Reducing Mitochondrial ROS

doi: 10.3390/antiox15030357

Figure Lengend Snippet: 2-methoxystypandrone (2-MS) inhibits mitochondrial ROS production in senescent fibroblasts. ( A ) Senescent fibroblasts treated with DMSO (0.01%), emodin-1-O-β-glucoside (8 μM), emodin-8-glucoside (8 μM), 2-methoxystypandrone (2-MS) (8 μM), or polydatin (8 μM) for 12 days at 4-day intervals. Mitochondrial ROS were measured after staining cells with 30 µM dihydrorhodamine 123 (DHR123. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s post–hoc test, with results considered not significant (n.s.) or significant at ** p < 0.01. Data represent the mean ± S.D., n = 3. ( B ) Senescent fibroblasts treated with DMSO (0.01%) or 2-MS (4, 8, and 12 μM) for 12 days at 4-day intervals. Mitochondrial ROS were measured after staining cells with 30 µM DHR123. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s post–hoc test, with results considered not significant (n.s.) or significant at ** p < 0.01. Data represent the mean ± S.D., n = 3. ( C ) Senescent fibroblasts treated with DMSO (0.01%) or 2-MS (4, 8, and 12 μM) for 12 days at 4-day intervals. DNA tail length was measured using the neutral comet assay. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s post–hoc test, with results considered significant at ** p < 0.01. For each condition, the experiment was repeated three times, yielding three independent sets of data (biological triplicates). Sixteen different samples were analyzed to generate each biological triplicate. Data represent the mean ± S.D.

Article Snippet: Mitochondrial ROS levels were measured in senescent fibroblasts treated with DMSO (D8418; Sigma, St. Louis, MO, USA, 0.01%), emodin-1-O-β-glucoside ( CFN93238 ; ChemFaces, Wuhan, China, 8 μM), emodin-8-glucoside (HY-N0311; MedChemExpress, Monmouth, NJ, USA, 8 μM), 2-MS ( CFN97412 ; ChemFaces, 8 μM), or polydatin (15721-25G; Sigma, 8 μM) for 12 days at 4-day intervals.

Techniques: Staining, Neutral Comet Assay