Journal: Journal of Diabetes Research

Article Title: Islet Transplantation Attenuating Testicular Injury in Type 1 Diabetic Rats Is Associated with Suppression of Oxidative Stress and Inflammation via Nrf-2/HO-1 and NF- κ B Pathways

doi: 10.1155/2019/8712492

Figure Lengend Snippet: Islet transplantation treatment inhibited NF- κ B expression in diabetic rats. (a, b) NF- κ B levels measured by immunohistochemical staining (×400) (bar = 25 μ m) and quantitative analysis of positive regions showed that the increase in NF- κ B was significantly improved in the INS group compared with that in the control group; IT attenuated the expression of the protein ( n = 6 for each group). (c, d) Western blot images of NF- κ B and quantitative analysis of its expression levels in the DM group revealed significant upregulation. By contrast, NF- κ B in the IT group was significantly downregulated compared with that in the INS group ( n = 6 for each group). (e–h) Typical Western blot images of TNF- α , IL-1 β , and IL-6 intesticular tissues and quantitative analysis of their expression levels. DM caused a significant increase in the above parameters. The IT treatment group significantly reduced the expression of TNF- α , IL-1 β , and IL-6. The expression of the above proteins in the INS group was only slightly altered ( n = 6 for each group). DM: diabetes mellitus; INS: insulin treatment; IT: islet transplantation. ∗ P < 0.05 vs. control; & P < 0.05 vs. DM; + P < 0.05 vs. INS.

Article Snippet: The membranes were blocked with 5% fat-free milk for 2 h at room temperature, followed by incubation with the following antibodies at 4°C overnight: nuclear factor like-2 factor (Nrf2; Abcam 1 : 1,000), HO-1 (Abcam 1 : 1,000), quinone oxidoreductase-1 (NQO-1; Abcam 1 : 1,000), NF- κ B (CST 1 : 1,000), necrosis factor alpha (TNF- α ) (CST 1 : 1,000), interleukin-1 beta (IL-1 β ) (CST 1 : 1,000), interleukin-6 (IL-6) (CST 1 : 1,000), and beta-actin (Bioworld 1 : 10,000).

Techniques: Transplantation Assay, Expressing, Immunohistochemical staining, Staining, Western Blot

Journal: Drug Design, Development and Therapy

Article Title: Isoliquiritigenin Attenuates UUO-Induced Renal Inflammation and Fibrosis by Inhibiting Mincle/Syk/NF-Kappa B Signaling Pathway

doi: 10.2147/DDDT.S243420

Figure Lengend Snippet: Isoliquiritigenin reduced the inflammatory response induced by LPS in BMDM. ( A – D ) ISL can reduce the mRNA expression of inflammatory cytokines in LPS-induced BMDM, including IL-1β, IL-6, TNF-α and MCP-1. ** P <0.01, vs LPS group. *** P <0.001, vs LPS group. **** P <0.0001, vs LPS group; ( E, F ) ISL reduced the secretion of IL-1β and IL-6 in supernatant of LPS-stimulated BMDM. ** P <0.01, vs LPS group; ( G ) immunofluorescence results showed that ISL inhibited the protein level of IL-1β and IL-6 in LPS-stimulated BMDM.

Article Snippet: Subsequently, the cells were treated with 0.5% Triton X-100 for 10 min at room temperature to increase the permeation of cell membrane, followed by blocking with 5% BSA for 30 min and then incubated with mouse anti- IL-1β antibody (1:200, Santa Cruz) or mouse anti-IL-6 antibody (ab9324, 1:150, CST) overnight at 4 °C.

Techniques: Expressing, Immunofluorescence

Journal: Drug Design, Development and Therapy

Article Title: Isoliquiritigenin Attenuates UUO-Induced Renal Inflammation and Fibrosis by Inhibiting Mincle/Syk/NF-Kappa B Signaling Pathway

doi: 10.2147/DDDT.S243420

Figure Lengend Snippet: Isoliquiritigenin protects the kidney by inhibiting Mincle/Syk/NF-kappa B signaling pathway and reducing the expression and secretion of inflammatory cytokines in the kidney and blood of UUO model. ( A ) Immunohistochemistry results showed that the infiltration of macrophage was significantly increased in the kidney of UUO model and ISL reduced infiltrated macrophage; ( B – F ) immunofluorescence and Western blot results showed that ISL can effectively reduce the protein level of Mincle and iNOS as well as phosphorylated Syk and NF-kappa B in the kidney of UUO model; ( G – J ) ISL strongly reduced the mRNA expression of IL-1β, IL-6, TNF-α and MCP-1 in the kidney of UUO model; ( K, L ) ISL also inhibited the secretion of IL-1β and IL-6 in the serum of UUO mice. Notes: * P <0.05, vs UUO group. ** P <0.01, vs UUO group. *** P <0.001 vs UUO group.

Article Snippet: Subsequently, the cells were treated with 0.5% Triton X-100 for 10 min at room temperature to increase the permeation of cell membrane, followed by blocking with 5% BSA for 30 min and then incubated with mouse anti- IL-1β antibody (1:200, Santa Cruz) or mouse anti-IL-6 antibody (ab9324, 1:150, CST) overnight at 4 °C.

Techniques: Expressing, Immunohistochemistry, Immunofluorescence, Western Blot

Journal: Drug Design, Development and Therapy

Article Title: Isoliquiritigenin Attenuates UUO-Induced Renal Inflammation and Fibrosis by Inhibiting Mincle/Syk/NF-Kappa B Signaling Pathway

doi: 10.2147/DDDT.S243420

Figure Lengend Snippet: Isoliquiritigenin suppressed inflammation through Mincle-dependent mechanism in BMDM cells. ( A ) In the Mincle recovery experiment, we used TDB to upregulate Mincle in ISL-treated inflammatory BMDM. The results showed that the protein level of Mincle ( B ), iNOS ( C ), phosphorylated Syk ( D ) and phosphorylated NF-kappa B ( E ) were significantly increased in ISL-treated inflammatory BMDM, suggesting ISL inhibits inflammation by suppressing Mincle; Overexpression of Mincle by administration of TDB, upregulated the expression ( F–I ) and secretion ( J–L ) of IL-1β, IL-6 and TNF-α and MCP-1 in ISL-treated inflammatory BMDM. Notes: * P <0.05, vs LPS group. ** P <0.01, vs LPS group. *** P <0.001, vs LPS group. # P <0.05, vs LPS+ISL 40μM-treated group. ## P <0.01, vs LPS+ISL 40μM-treated group. ### P <0.001, vs LPS+ISL 40μM-treated group.

Article Snippet: Subsequently, the cells were treated with 0.5% Triton X-100 for 10 min at room temperature to increase the permeation of cell membrane, followed by blocking with 5% BSA for 30 min and then incubated with mouse anti- IL-1β antibody (1:200, Santa Cruz) or mouse anti-IL-6 antibody (ab9324, 1:150, CST) overnight at 4 °C.

Techniques: Over Expression, Expressing

Journal: Molecular Therapy Oncolytics

Article Title: Dual Inhibition of MAPK and JAK2/STAT3 Pathways Is Critical for the Treatment of BRAF Mutant Melanoma

doi: 10.1016/j.omto.2020.06.004

Figure Lengend Snippet: Crosstalk between the JAK2/STAT3 and MAPK Pathways in A375 and A375R Cells (A and B) A375 and A375R cells were treated with PLX4032 (A), PLX4032 and WP1066 (B) for 6 h. Phospho-STAT3 (705), phospho-STAT3 (727), STAT3, phospho-ERK1/2, ERK, and IL-6 (A only) levels were analyzed by western blotting, and tubulin served as a loading control. (C) A375 and A375R cells were treated with different concentrations of WP1066 or PLX4032 and WP1066 (2 μM) for 72 h. Cell viability was determined by CCK8 assays. Data are mean ± SD. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; Student’s t test.

Article Snippet: Western blotting was performed using the following antibodies: anti-IL-6 (1:1,000; CST, MA, USA), anti-ERK (1:2,000; CST, MA, USA), anti-phospho-ERK (1:2,000; CST, MA, USA), anti-phospho-STAT3 (705; 1:1,000; CST, MA, USA), anti-phospho-STAT3 (727; 1:1,000; CST, MA, USA), anti-STAT3 (1:1,000; CST, MA, USA), anti-α-tubulin (1:2,000; ABclonal, Wuhan, China), and anti-VEGFA (1:1,000; BioWorld, Nanjing, China).

Techniques: Western Blot