Fig 1: Effects of yeast extract on wrinkle-related protein expression in HDF. Collagen-1 (A) and elastin (B) protein levels were significantly reduced after UVB irradiation, and yeast extract reversed this effect. On the other hand, UVB exposure enhanced the protein expression of MMP-1 (C), MMP-3 (D), and MMP-9 (E), but yeast extract successfully countered these influences. MMP, matrix metalloproteinase; RA, Retinoic acid, 1 µM; UVB, ultraviolet B. ****p < 0.001, independent samples t-test compared to non-irradiated control group; ###p < 0.005, ####p < 0.001, independent samples t-test compared to UVB-irradiated control group
Fig 2: The decreased USP26 expression is correlated with aging of BMSCs. A) Representative H&E staining images of the femurs from 2‐month‐old, 15‐month‐old, and 20‐month‐old mice. n = 5 in each group. Scale bar, 500 µm. B) Representative Micro‐CT images of the femurs from 2‐month‐old, 15‐month‐old, and 20‐month‐old mice. n = 5 in each group. C) Quantitative analysis of the trabecular bone from 2‐month‐old, 15‐month‐old, and 20‐month‐old mice including BV/TV, Tb.N, Tb.Th, Tb.Sp, and BMD. n = 5 in each group. D) Western‐blot analysis of USP26, P21, and P16 protein levels in BMSCs from mice of different ages (2, 15, and 20 months). n = 3 in each group. E) qPCR analysis of Usp26, P16, and P21 mRNA expressions in BMSCs from mice of different ages (2, 15, and 20 months). n = 3 in each group. F) Western blot analysis of USP26, P21, and P16 protein levels in BMSCs of different generations (1, 5, 10, and 15). n = 3 in each group. G) qPCR analysis of Usp26 mRNA expressions from the bone marrow of different patients (n = 15) and evaluation of the relevance between Usp26 mRNA expression and age. H) Representative SA‐β‐Gal staining images and percentage of SA‐β‐Gal positive cells of BMSCs from control and cKO mice. n = 5 in each group. Scale bar, 10 µm. I) Western blot analysis of USP26, P21, and P16 protein levels of BMSCs from control and cKO mice. n = 3 in each group. J) qPCR analysis of Usp26, P16, and P21 mRNA expressions of BMSCs from control and cKO mice. n = 3 in each group. K) qPCR analysis of SASP‐related gene (Il‐1α, Il‐1β, Il‐6, Cxcl1, Cxcl10, Ccl2, and Ccl5) mRNA expressions of BMSCs from control and cKO mice. n = 3 in each group. L) Representative Immunohistochemistry (IHC) staining of IL‐6 of the femurs from control and cKO mice. IL‐6 positive cells number as a quantitative measurement. n = 6 in each group. Scale bar, 50 µm (black) and 10 µm (red). M) Total protein lysates of bone marrow supernatant from the femurs of control and cKO mice were analyzed for TNF‐α, IL‐1α, IL‐6, MMP3, and MMP13. n = 6 in each group. N) Representative immunofluorescent images of γ‐H2AX foci in BMSCs from control and cKO mice and quantification of the number of γ‐H2AX foci per cell. n = 6 in each group. Mice age in H), I), J), K), L), M), and N), 2‐month‐old. BMSCs from mice were used in D), E), F), H), I), J), K), M), and N); BMSCs from Human were used in G). Data are represented as mean ± SD. Statistical significance was determined by one‐way ANOVA in C) and E), and two‐sided student's t test in H), J), L), M) and N). *p < 0.05, **p < 0.01, ***p < 0.001.
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