Fig 2: (A) Western blot analysis of the protein expression of the fibroblast markers PDGFR-a, PDGFR-ß, and a-SMA, and the mesenchymal marker CD45 in heart cells obtained from parental wild-type (WT) and Gpx-1 knockout (KO) animals. (B) Northern analysis of Gpx-1 expression in cardiac fibroblasts from parental (+) and Gpx-1 knockout (-) C57/Bl6 mice. Cells were passaged without (-/-; +/-) or with (-/+; +/+) supplemental sodium selenite (100 nM). The presence of exogenous selenium increased Gpx-1 mRNA expression by ˜ 3-fold.

Fig 3: Silencing p53 enhances vasculogenic differentiation of DPSC in vitro and in vivo.a Photomicrographs depicting dental pulp stem cells that differentiated into functional blood vessels in vivo. GFP-transduced DPSC were seeded in scaffolds and transplanted in the subcutaneous space of the dorsum of SCID mice. Six weeks later, scaffolds were retrieved and tissue sections were prepared for immunofluorescence staining. Scattered cells showed positive staining for both, GFP (green), and SMA-alpha (red). In mature blood vessels, GFP was detected in the endothelial cells, SMA-alpha was stained the surrounding smooth muscle cells. Endothelial cells in the inner layer were stained for CD31 (green), while surrounding smooth muscle cells stained positive for SMA-alpha (red). Arrows point to DPSC-derived endothelial cells (green cells). Scale bar: 50 µm. b Western blots for VEGFR2, CD31, PDGFR-a, PDGFR-ß, SMA-a, p53, and Bmi-1 in DPSC, SHED, HUASMC, and HDMEC. c Endothelial cell differentiation: shRNA-transduced DPSC were cultured with 5%FBS–MEM for 14 days, western blots for p53, p21, VEGFR2, Tie-2, CD31, and VE-cadherin. The density of protein expression was normalized with ß-actin. d shRNA-p53-mediated vasculogenic differentiation in vivo: H&E and IHC staining. Blood vessels were revealed by H&E staining and detected with anti-factor VIII antibody, scale bar: 50 µm. e Graph depicting the blood vessel density observed in d stained with Factor VIII. Asterisk indicates p = 0.00035, as determined by t test. f DPSC cells were seeded in matrigel and cultured with EGM2 for 8 days. The Matrigel was fixed, and the sprouts were revealed by IF staining for CD31. Scale bar: 100 µm. g In all, 1 × 104 shRNA-transduced DPSC were seeded in growth factor-reduced matrigel-coated 12-well plate and cultured in endothelial differentiation medium (EGM2) for indicated time points. Sprouts were photographed, scale bar:100 µm. h Graph depicting the numbers of sprout formed in g. Three independent experiments using triplicate wells per condition were performed. Asterisk indicates p < 0.001, as determined by one-way ANOVA followed by a post hoc test (Tukey’s test).

Fig 4: mRNA and protein levels of PDGF receptor-ß produced by chondrocytes in superficial and deep zones. PDGFRB mRNA level in superficial-zone chondrocytes (SZC) was 4 times higher than that in deep-zone chondrocytes (DZC), as assessed by quantitative reverse transcription-PCR (?Ct method) (A). The protein level of PDGF receptor-ß was > 4 times higher in the superficial zone than in the deep zone, as assessed by Western blot analysis (B). The blots were cropped from different parts of the same gel; and full-length gels and blots are included in the Supplementary Information. **p < 0.01, Mann-Whitney U-test

Fig 5: Effect of inhibition of Glo-I by BrBzGSHCp2 on proliferation and related signaling pathways in Huh7 cells. (A) Specific activity of Glo-I was significantly reduced after 24 h of BrBzGSHCp2 treatment in a concentration-dependent manner indicating strong inhibition of enzymatic activity. (B) Huh7 cells were incubated at different time points (6, 24 h) with increasing levels of BrBzGSHCp2 (1–10 µM). WST assays revealed significant dose-dependent reduction of cell proliferation at each time points. (C1–C7) Huh7 cells were incubated for 24 h with 1–10 µM BrBzGSHCp2. Protein analysis showed significant influence on proliferation signaling pathways (reduction of PDGFR-ß, VEGFR2, VEGF, pERK/ERK, and NF-?B; increase of Nrf2). Representative Western Blot images are shown in (C1) quantifications (C2–C7) are expressed as mean ± S.D. of at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.