Journal: PLOS ONE

Article Title: MiR-132-3p suppresses peritoneal fibrosis induced by peritoneal dialysis via targeting TGF- β 1/Smad2/3 signaling pathway

doi: 10.1371/journal.pone.0301540

Figure Lengend Snippet: Sequences of the primers for q-PCR.

Article Snippet: The primary antibodies used were as follows: Smad2 (A00090-1, Boster, 1:200 dilution; China); Smad3 (BM3919, Boster, 1:200 dilution; China); α-SMA (BM0002, Boster, 1:200 dilution; China); TGF- β 1 (MA00019, Boster, 1:200 dilution; China).

Techniques:

Journal: PLOS ONE

Article Title: MiR-132-3p suppresses peritoneal fibrosis induced by peritoneal dialysis via targeting TGF- β 1/Smad2/3 signaling pathway

doi: 10.1371/journal.pone.0301540

Figure Lengend Snippet: (A) Area semi-quantitative statistical analysis by immunohistochemistry; (B-E)Expression of TGF- β 1, smad2, smad3,col1a1 col3a1and α-SMA by immunohistochemistry. All data are presented as mean ± SD. n = 6 for each group. * p < 0.05; ** p < 0.01; *** p < 0.001; between the indicated groups.

Article Snippet: The primary antibodies used were as follows: Smad2 (A00090-1, Boster, 1:200 dilution; China); Smad3 (BM3919, Boster, 1:200 dilution; China); α-SMA (BM0002, Boster, 1:200 dilution; China); TGF- β 1 (MA00019, Boster, 1:200 dilution; China).

Techniques: Immunohistochemistry, Expressing

Journal: PLOS ONE

Article Title: MiR-132-3p suppresses peritoneal fibrosis induced by peritoneal dialysis via targeting TGF- β 1/Smad2/3 signaling pathway

doi: 10.1371/journal.pone.0301540

Figure Lengend Snippet: Quantitative real time-PCR was used to measure the mRNA levels of(A)TGF- β1 ;(B)α-SMA;(C)Smad2;(D)Smad3. All data are presented as mean ± SD. n = 4 for each group. * p < 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001 between the indicated groups.

Article Snippet: The primary antibodies used were as follows: Smad2 (A00090-1, Boster, 1:200 dilution; China); Smad3 (BM3919, Boster, 1:200 dilution; China); α-SMA (BM0002, Boster, 1:200 dilution; China); TGF- β 1 (MA00019, Boster, 1:200 dilution; China).

Techniques: Real-time Polymerase Chain Reaction

Journal: PLOS ONE

Article Title: MiR-132-3p suppresses peritoneal fibrosis induced by peritoneal dialysis via targeting TGF- β 1/Smad2/3 signaling pathway

doi: 10.1371/journal.pone.0301540

Figure Lengend Snippet: (A and D) Immunofluorescence was used to measure the protein expression levels of(B)TGF- β 1;(C)Smad3;(E) α -SMA;(F)Smad2. All data are presented as mean ± SD. n = 3 for each group. * p < 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001 between the indicated groups.

Article Snippet: The primary antibodies used were as follows: Smad2 (A00090-1, Boster, 1:200 dilution; China); Smad3 (BM3919, Boster, 1:200 dilution; China); α-SMA (BM0002, Boster, 1:200 dilution; China); TGF- β 1 (MA00019, Boster, 1:200 dilution; China).

Techniques: Immunofluorescence, Expressing

Journal: JCI Insight

Article Title: TGF- β promotes fibrosis after severe acute kidney injury by enhancing renal macrophage infiltration

doi: 10.1172/jci.insight.123563

Figure Lengend Snippet: Peritoneal macrophages from Tgfbr2fl/fl (WT) and LysM-Cre Tgfbr2fl/fl (macrophage TGF-βRII–/–; MφTGF-βRII–/–) mice 3 days after peritoneal thioglycollate administration were used. (A) TGF-βRII levels were markedly lower in TGF-βRII–/– macrophages than in WT macrophages. ***P < 0.001, n = 3 in each group. Data were expressed as mean ± SEM. (B) Macrophages were stimulated with 2 ng/ml recombinant TGF-β for 30 minutes. TGF-βRII deletion led to decreases in TGF-β–stimulated phosphorylation of Smad2 and Smad3, an indication of TGF-βRII deficiency. p-Smad3, phospho-Smad3; p-Smad2, phospho-Smad2; FAK, focal adhesion kinase.

Article Snippet: Rat anti-mouse F4/80 (MCA497R, a marker of macrophages) and CD3 (MCA1477, a marker of T lymphocytes) were purchased from AbD Serotec (now Bio-Rad); mouse anti–mannose receptor (CD206, MAB25341) and mouse anti–4-HNE (a marker of oxidative stress, 198960) were from R&D Systems; rabbit anti–TGF-βRII (SC-400) and goat anti-human CTGF (SC-14939) were from Santa Cruz Biotechnology; rabbit anti-human Smad2 (600-401-A59), phospho-Smad2 (600-401-K09S), and phospho-Smad3 (600-401-919) as well as rabbit anti-murine collagen type I (600-401-103-01) were from Rockland Immunochemicals; mouse anti–α-SMA (A5228, a marker of myofibroblasts) was from Sigma-Aldrich; and rabbit anti–TGF-β1 (NBP1-80289) and mouse anti–TGF-β2 (Mab612-SP) were from Novus Biologicals.

Techniques: Recombinant

Journal: Molecular Medicine Reports

Article Title: Astaxanthin ameliorates renal interstitial fibrosis and peritubular capillary rarefaction in unilateral ureteral obstruction

doi: 10.3892/mmr.2019.9970

Figure Lengend Snippet: ASX inhibits UUO-induced activation of the TGF-β1/Smad signaling pathway. Protein expression levels of (A) TGF-β1, (B) Smad2, (C) Smad3, (D) Smad4 and (E) Smad7 in murine renal tissues were assessed by western blotting. The data are presented as the means ± standard deviation (n=6). **P<0.01, ***P<0.001. ASX, Astaxanthin; p, phosphorylated; TGF-β1, transforming growth factor β1; UUO, unilateral ureteral obstruction.

Article Snippet: After blocking with 5% skimmed milk for 1 h at room temperature, the membranes were probed with primary antibodies against collagen I (1:2,000; cat. no. 14695-1-AP; ProteinTech Group, Inc.), TSP-1 (1:500; cat. no. 18304-1-AP; ProteinTech Group, Inc.), VEGF-A (1:1,000; cat. no. 19003-1-AP; ProteinTech Group, Inc.), TGF-β1 (1:500; cat. no. 21898-1-AP; ProteinTech Group, Inc.), phosphorylated (p)-Smad2 (1:1,000; cat. no. 3108; Cell Signaling Technology, Inc., Danvers, MA, USA), Smad2 (1:3,000; cat. no. 12570-1-AP; ProteinTech Group, Inc.), fibronectin (1:1,000; cat. no. 15613-1-AP; ProteinTech Group, Inc.), α-smooth muscle actin (α-SMA) (1:500; cat. no. 55135-1-AP; ProteinTech Group, Inc.), Smad3 (1:1,000; cat. no. bs-3484R; BIOSS, Beijing, China), p-Smad3 (1:1,000; cat. no. bsm-52205R; BIOSS), Smad4 (1:1,000; cat. no. bs-0585R; BIOSS), Smad7 (1:1,000; cat. no. bs-23328R; BIOSS) and β-actin (1:500; cat. no. bsm-33036M; BIOSS) overnight at 4°C.

Techniques: Activation Assay, Expressing, Western Blot, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Astaxanthin ameliorates renal interstitial fibrosis and peritubular capillary rarefaction in unilateral ureteral obstruction

doi: 10.3892/mmr.2019.9970

Figure Lengend Snippet: ASX suppresses TGF-β1-induced expression of profibrogenic factors via inactivation of the Smad signaling pathway. Protein expression levels of (A) collagen I, (B) α-SMA, (C) Smad2, (D) Smad3, (E) Smad4 and (F) Smad7 in NRK-52E cells were assessed by western blotting. The data are presented as the means ± standard deviation (n=6). *P<0.05, **P<0.01, ***P<0.001. ASX, Astaxanthin; NS, not significant; p, phosphorylated; TGF-β1, transforming growth factor β1; UUO, unilateral ureteral obstruction.

Article Snippet: After blocking with 5% skimmed milk for 1 h at room temperature, the membranes were probed with primary antibodies against collagen I (1:2,000; cat. no. 14695-1-AP; ProteinTech Group, Inc.), TSP-1 (1:500; cat. no. 18304-1-AP; ProteinTech Group, Inc.), VEGF-A (1:1,000; cat. no. 19003-1-AP; ProteinTech Group, Inc.), TGF-β1 (1:500; cat. no. 21898-1-AP; ProteinTech Group, Inc.), phosphorylated (p)-Smad2 (1:1,000; cat. no. 3108; Cell Signaling Technology, Inc., Danvers, MA, USA), Smad2 (1:3,000; cat. no. 12570-1-AP; ProteinTech Group, Inc.), fibronectin (1:1,000; cat. no. 15613-1-AP; ProteinTech Group, Inc.), α-smooth muscle actin (α-SMA) (1:500; cat. no. 55135-1-AP; ProteinTech Group, Inc.), Smad3 (1:1,000; cat. no. bs-3484R; BIOSS, Beijing, China), p-Smad3 (1:1,000; cat. no. bsm-52205R; BIOSS), Smad4 (1:1,000; cat. no. bs-0585R; BIOSS), Smad7 (1:1,000; cat. no. bs-23328R; BIOSS) and β-actin (1:500; cat. no. bsm-33036M; BIOSS) overnight at 4°C.

Techniques: Expressing, Western Blot, Standard Deviation