Fig 1: Schematic diagram of the mechanism of action of ASB in UV-induced photo-damage in HaCaT cells. UV, ultraviolet; ASB, androgra-pholide sodium bisulfite; ROS, reactive oxygen species; keap1, kelch-like ECH-associated protein 1; Nrf2, nuclear factor E2-related factor 2; IL, interleukin; TNF-a, tumor necrosis factor-a; GCLC, glutamate-cysteine ligase catalytic subunit; NQO1, NAD(P)H quinone oxidoreductase 1; I?Ba, NF-?B inhibitor-a; ARE, antioxidant response element.
Fig 2: The expression of p62, Keap1 and Nrf2. (A and B) Representative images and (C) quantification revealing the levels of p62 in (A) BPH and (B) PCa. A total of 150 patients including 112 patients with PCa and 38 with BPH were evaluated. The relative levels of p62 in prostate glands and stromas and in cytoplasm and nuclei were compared between BPH and PCa. *P=0.05 and **P=0.01 Representative images showing (D) high levels of Keap1 in BPH, or (E) low levels of Keap1 in PCa, and (F) low levels of Nrf2 in BPH, or (G) high levels of Nrf2 in PCa. Representative images showing (H) low or (I and J) high levels of p62 expressed in prostate tissues from (H) normal, (I) PTEN-/-, or (J) TRAMP mice; (K) high, or (L and M) low levels of Keap1 expressed in (K) prostate tissues from normal, (L) PTEN-/-, or (M) TRAMP mice; and (N) low, or (O and P) high levels of Nrf2 expressed in (N) prostate tissues from normal, (O) PTEN-/-or (P) TRAMP mice. Normal prostate tissues and prostate tumors were collected from three sets of wild-type, PTEN-/- and TRAMP mice. Scale bar, 50 µm. Keap1, Kelch-like ECH-associated protein 1; Nrf2, nuclear factor erytheroid-derived 2-like 2; BPH, benign prostate hyperplasia; PCa, prostate cancer; TRAMP, transgenic adenocarcinoma of mouse prostate.
Fig 3: p62 suppresses the expression of Keap1 and enhances the activity of Nrf2. C1, DU145 cells carrying the control plasmid vector; OE, DU145 cells overexpressing p62; KO, OE cells with p62 knocked out by CRISPER technology; and C2, OE cells carrying negative control for CRISPR knockout. (A-P) Representative (A, C, E, G, I, K, M and O) immunoblots and (B, D, F, H, J, L, N and P) quantification showing levels of (A, B, I and J) p62, (C, D, K and L) Keap1, and (E, F, M and N) total and (G, H, O and P) nuclear Nrf2 in (A-H) C1 and OE cells and (I-P) C2 and KO cells. (Q-T) Representative images revealing the (Q and R) immunostaining intensities and (S and T) quantification of (Q and S) Keap1 and (R and T) Nrf2 in C1, OE, C2 and KO cells. The nuclei were stained with DAPI (blue). All images were captured under an identical setting. The results represent three independent experiments. *P=0.05, **P=0.01, and ***P=0.001. Keap1, Kelch-like ECH-associated protein 1; Nrf2, nuclear factor erytheroid-derived 2-like 2.
Fig 4: Changes in the Nrf2 and NF-?B signaling pathways in HaCaT cells after UV irradiation. (A) Nuclear and cytoplasmic proteins were extracted from the cultured cells at 0, 0.5, 1, 2, 3, 4, 5 and 6 h after UV irradiation (90 mJ/cm2). The protein expression levels of NF-?B-mediated p65, and I?Ba and Nrf2-mediated Nrf2 and keap1, were measured by western blotting. (B) Relative changes in protein intensities were quantified by densitometric analysis and are presented as bar diagrams (n=3 for each group). The results are expressed as the mean ± SD. aP<0.05 vs. the 0 h group; bP<0.05 vs. the 0.5 h group; cP<0.05 vs. the 1 h group; dP<0.05 vs. the 2 h group; eP<0.05 vs. the 3 h group; fP<0.05 vs. the 4 h group; gP<0.05 vs. the 5 h group. UV, ultraviolet; I?Ba, NF-?B inhibitor-a; keap1, kelch-like ECH-associated protein 1; Nrf2, nuclear factor E2-related factor 2.
Fig 5: Effect of piceatannol treatment on Western blotting results of cytosolic Nrf2, nuclear Nrf2, Keap1, NQO1, HO1, ?GCS, and GPx in the testes of control and cadmium-treated rats.Abbreviations: C, control; PT, piceatannol (10 mg/kg); Cd, cadmium (5 mg/kg).
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