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( A ) Western blot results for the detection of pSmad5, pSmad1/5/9, and other proteins after enalapril treatment. ( B ) GSEA showing the enrichment of the BMP signaling pathway after enalapril treatment. NES, normalized enrichment score. ES, enrichment score. ( C ) Profile of pSmad1/5/9 enrichment at the TSS regions in the control group (Ctrl) and the enalapril-treated group (Enalapril). ( D ) Heatmap of cell cycle (green) and SASP-related genes (purple) in the control group (Ctrl), BMP receptor inhibitor-treated group (LDN193189, LDN), and enalapril and BMP receptor inhibitor-cotreated group (EP+LDN). ( E ) Western blot analysis showing the protein levels of pSmad1/5/9, p16, and p21 in response to different combinations of enalapril, a BMP receptor inhibitor (LDN193189, LDN), and <t>BMP4.</t> ( F ) Senescence-associated β-galactosidase (SA-β-Gal) staining (left) and corresponding ratio statistical chart (right) of IMR90 cells treated with different combinations of enalapril, a BMP receptor inhibitor (LDN193189, LDN), and BMP4. Scale bars, 200 μm. Enlarged scale bars, 100 μm. ( G ) Ki67 immunofluorescence experiment (left) and intensity statistical chart (right) of IMR90 cells treated with different combinations of enalapril, a BMP receptor inhibitor (LDN193189, LDN), and BMP4. Scale bars, 80 μm. Enlarged scale bars, 40 μm. A two-tailed t-test was employed; ns indicates no significant difference, * p <0.05, ** p <0.01, *** p <0.001. Figure 2—source data 1. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 2—source data 2. Original files for western blot analysis displayed in .
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( A ) Western blot results for the detection of pSmad5, pSmad1/5/9, and other proteins after enalapril treatment. ( B ) GSEA showing the enrichment of the BMP signaling pathway after enalapril treatment. NES, normalized enrichment score. ES, enrichment score. ( C ) Profile of pSmad1/5/9 enrichment at the TSS regions in the control group (Ctrl) and the enalapril-treated group (Enalapril). ( D ) Heatmap of cell cycle (green) and SASP-related genes (purple) in the control group (Ctrl), BMP receptor inhibitor-treated group (LDN193189, LDN), and enalapril and BMP receptor inhibitor-cotreated group (EP+LDN). ( E ) Western blot analysis showing the protein levels of pSmad1/5/9, p16, and p21 in response to different combinations of enalapril, a BMP receptor inhibitor (LDN193189, LDN), and BMP4. ( F ) Senescence-associated β-galactosidase (SA-β-Gal) staining (left) and corresponding ratio statistical chart (right) of IMR90 cells treated with different combinations of enalapril, a BMP receptor inhibitor (LDN193189, LDN), and BMP4. Scale bars, 200 μm. Enlarged scale bars, 100 μm. ( G ) Ki67 immunofluorescence experiment (left) and intensity statistical chart (right) of IMR90 cells treated with different combinations of enalapril, a BMP receptor inhibitor (LDN193189, LDN), and BMP4. Scale bars, 80 μm. Enlarged scale bars, 40 μm. A two-tailed t-test was employed; ns indicates no significant difference, * p <0.05, ** p <0.01, *** p <0.001. Figure 2—source data 1. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 2—source data 2. Original files for western blot analysis displayed in .

Journal: eLife

Article Title: Enalapril mitigates senescence and aging-related phenotypes in human cells and mice via pSmad1/5/9-driven antioxidative genes

doi: 10.7554/eLife.104774

Figure Lengend Snippet: ( A ) Western blot results for the detection of pSmad5, pSmad1/5/9, and other proteins after enalapril treatment. ( B ) GSEA showing the enrichment of the BMP signaling pathway after enalapril treatment. NES, normalized enrichment score. ES, enrichment score. ( C ) Profile of pSmad1/5/9 enrichment at the TSS regions in the control group (Ctrl) and the enalapril-treated group (Enalapril). ( D ) Heatmap of cell cycle (green) and SASP-related genes (purple) in the control group (Ctrl), BMP receptor inhibitor-treated group (LDN193189, LDN), and enalapril and BMP receptor inhibitor-cotreated group (EP+LDN). ( E ) Western blot analysis showing the protein levels of pSmad1/5/9, p16, and p21 in response to different combinations of enalapril, a BMP receptor inhibitor (LDN193189, LDN), and BMP4. ( F ) Senescence-associated β-galactosidase (SA-β-Gal) staining (left) and corresponding ratio statistical chart (right) of IMR90 cells treated with different combinations of enalapril, a BMP receptor inhibitor (LDN193189, LDN), and BMP4. Scale bars, 200 μm. Enlarged scale bars, 100 μm. ( G ) Ki67 immunofluorescence experiment (left) and intensity statistical chart (right) of IMR90 cells treated with different combinations of enalapril, a BMP receptor inhibitor (LDN193189, LDN), and BMP4. Scale bars, 80 μm. Enlarged scale bars, 40 μm. A two-tailed t-test was employed; ns indicates no significant difference, * p <0.05, ** p <0.01, *** p <0.001. Figure 2—source data 1. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 2—source data 2. Original files for western blot analysis displayed in .

Article Snippet: Recombinant BMP4 protein (MCE, HY-P7007) was dissolved in sterile ddH 2 O to prepare a 100 μg/mL stock solution.

Techniques: Western Blot, Control, Staining, Immunofluorescence, Two Tailed Test

( A ) Western blot analysis showing the changes in protein levels following BMP4 treatment. ( B ) Senescence-associated β-galactosidase (SA-β-Gal) staining (left) and corresponding ratio analysis chart (right) following BMP4 treatment. Scale bars, 200 μm. Enlarged scale bars, 100 μm. ( C ) Ki67 immunofluorescence (right) and corresponding ratio analysis chart (left) following BMP4 treatment. Scale bars, 80 μm. Enlarged scale bars, 40 μm. A two-tailed t-test was employed, ** p <0.01, **** p <0.0001. Figure 2—figure supplement 2—source data 1. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 2—figure supplement 2—source data 2. Original files for western blot analysis displayed in .

Journal: eLife

Article Title: Enalapril mitigates senescence and aging-related phenotypes in human cells and mice via pSmad1/5/9-driven antioxidative genes

doi: 10.7554/eLife.104774

Figure Lengend Snippet: ( A ) Western blot analysis showing the changes in protein levels following BMP4 treatment. ( B ) Senescence-associated β-galactosidase (SA-β-Gal) staining (left) and corresponding ratio analysis chart (right) following BMP4 treatment. Scale bars, 200 μm. Enlarged scale bars, 100 μm. ( C ) Ki67 immunofluorescence (right) and corresponding ratio analysis chart (left) following BMP4 treatment. Scale bars, 80 μm. Enlarged scale bars, 40 μm. A two-tailed t-test was employed, ** p <0.01, **** p <0.0001. Figure 2—figure supplement 2—source data 1. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 2—figure supplement 2—source data 2. Original files for western blot analysis displayed in .

Article Snippet: Recombinant BMP4 protein (MCE, HY-P7007) was dissolved in sterile ddH 2 O to prepare a 100 μg/mL stock solution.

Techniques: Western Blot, Staining, Immunofluorescence, Two Tailed Test