Journal: Acta Pharmaceutica Sinica. B

Article Title: USP25 ameliorates vascular remodeling by deubiquitinating FOXO3 and promoting autophagic degradation of FOXO3

doi: 10.1016/j.apsb.2024.12.033

Figure Lengend Snippet: Usp25 deletion promoting aortic EndMT in Ang II-challenged mice. (A, B) Murine aortic endothelial cells (MAECs) were exposed to 1 μmol/L Ang II for different time periods. Lysates were probed for USP25 (A). GAPDH was used as the loading control. Densitometric quantification of USP25 is shown in (B) (mean ± SEM; n = 3; ns = not significant, ∗ P < 0.05). (C, D) Murine aortic smooth muscle cells (MASMCs) were exposed to 1 μmol/L Ang II for different time periods. Lysates were probed for USP25 (C). GAPDH was used as the loading control. Densitometric quantification of USP25 is shown in (D) (mean ± SEM; n = 3; ns = not significant, ∗ P < 0.05). (E, F) Representative double immunofluorescence staining images showing expression of VIMENTIN (red) and CD31 (green) in panel (E). Sections were counterstained with DAPI (blue). Proportion of overlapping area is shown in (F) (Scale bar = 20 μm). (G, H) Representative double immunofluorescence staining images showing expression of SNAI1+SLUG (red) and CD31 (green) in panel (G). Sections were counterstained with DAPI (blue). Proportion of overlapping area is shown in (H) (Scale bar = 20 μm). (I, J) Representative double immunofluorescence staining images showing expression of VE-cadherin (red) and CD31 (green) in panel (I). Sections were counterstained with DAPI (blue). Proportion of overlapping area is shown in (J) (Scale bar = 20 μm). One-way ANOVA followed by Tukey's post hoc test for (B, D, F, H, J).

Article Snippet: Finally, anti-fluorescence quencher kit with DAPI (Cat# SP-8500-15; Vector Laboratories, USA) was used.

Techniques: Control, Double Immunofluorescence Staining, Expressing

Journal: Acta Pharmaceutica Sinica. B

Article Title: USP25 ameliorates vascular remodeling by deubiquitinating FOXO3 and promoting autophagic degradation of FOXO3

doi: 10.1016/j.apsb.2024.12.033

Figure Lengend Snippet: USP25 induction suppresses Ang II-induced vascular remodeling. Usp25 −/− mice were administered AAV9 expressing Usp25 . Empty AAV9 was administered in WT and Usp25 −/− mice as the control. Mice were then infused with Ang II for 4 weeks. (A, B) Representative low-power (A) and high-power (B) images of aortas stained with H&E (Scale bar = 200 μm in panel (A), 50 μm in panel (B); M = media). (C) Quantification of medial thickness in mice determined from H&E-stained sections (mean ± SEM; n = 6; ∗ P < 0.05). (D, E) Aortic tissues were stained with Masson's Trichrome. Representative staining images are shown in panel (D) (Scale bar = 50 μm; E = endothelium, A = adventitia). Quantification of collagen from Masson's Trichrome-stained sections is shown in panel (E) (mean ± SEM; n = 6; ∗ P < 0.05). (F, G) Representative immunohistochemical staining for TGF- β 1 (brown) in aortas are shown in panel (F) (Scale bar = 50 μm; E = endothelium, A = adventitia). Quantification of positive area from immunohistochemical sections of TGF- β 1 is shown in panel G (mean ± SEM; n = 6; ns = not significant, ∗ P < 0.05). (H, I) Representative double immunofluorescence staining images showing expression of VIMENTIN (red) and CD31 (green) in panel (H). Sections were counterstained with DAPI (blue). Proportion of overlapping area is shown in (I) (Scale bar = 20 μm). (J, K) Representative double immunofluorescence staining images showing expression of SNAI1+SLUG (red) and CD31 (green) in panel (J). Sections were counterstained with DAPI (blue). The proportion of overlapping area is shown in (K) (Scale bar = 20 μm). (L, M) Representative double immunofluorescence staining images showing expression of VE-cadherin (red) and CD31 (green) in panel (L). Sections were counterstained with DAPI (blue). The proportion of overlapping area is shown in (M) (Scale bar = 20 μm). One-way ANOVA followed by Tukey's post hoc test for (C, E, G, I, K, M).

Article Snippet: Finally, anti-fluorescence quencher kit with DAPI (Cat# SP-8500-15; Vector Laboratories, USA) was used.

Techniques: Expressing, Control, Staining, Immunohistochemical staining, Double Immunofluorescence Staining