Journal: Molecular Medicine Reports

Article Title: The protective role of vitamin D3 in a murine model of asthma via the suppression of TGF-β/Smad signaling and activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2016.5563

Figure Lengend Snippet: Effects of vitamin D3 on transforming growth factor-β (TGF-β)/Smad signaling in lungs induced by ovalbumin (OVA) challenge. (A) Levels of TGF-β1 in bronchoalveolar lavage fluid (BALF), as determined by enzyme-linked immunosorbent assay. (B) Representative protein expression of TGF-β1 determined by western blotting. (C) Relative protein levels of TGF-β1 in lung tissues. (D) Representative protein expression of phosphorylated (p)-Smad2, Smad2, p-Smad3 and Smad3 determined by western blotting. Relative protein levels of (E) p-Smad2 and (F) p-Smad3. Data are presented as the mean ± standard deviation of three repeat experiments, n=6. ** P<0.01 vs. the control group; ## P<0.01 vs. the OVA group.

Article Snippet: After blocking with 5% fat-free milk, the membranes were immunoblotted with primary antibodies as follows: Mouse α-SMA (1:400), rabbit HO-1 (1:200; Santa Cruz Biotechnology, Inc.; cat. no. sc-10789), mouse Nrf2 (1:400), rabbit TGF-β1 (1:200; Santa Cruz Biotechnology, Inc.; cat. no. sc-146), rabbit Smad2 (1:500, BIOSS, Beijing, China; cat. no. bs-0718R), rabbit phosphorylated (p)-Smad2 (1:500; BIOSS; cat. no. bs-5618R), rabbit Smad3 (1:500; BIOSS; cat. no. bs-3484R), rabbit p-Smad3 (1:500; BIOSS; cat. no. bs-5459R), mouse β-actin (1:1,000; Santa Cruz Biotechnology, Inc.; cat. no. sc-47778) and rabbit histone H3 (1:1,000; BIOSS; cat. no. bs-17422R) at 4°C overnight.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Standard Deviation

Journal: The Journal of pathology

Article Title: A 3D tri-culture system reveals that activin receptor-like kinase 5 and connective tissue growth factor drive human glomerulosclerosis

doi: 10.1002/path.4960

Figure Lengend Snippet: (A) MC 3D monoculture without the addition of TGFβ, and (B) in the presence of TGFβ, shown by H&E staining of paraffin-embedded sections. Human collagen type I/III (immunostained with an antibody that recognises both collagen type I and III) (C) and human collagen type IV (D) can be demonstrated in nodules by immunoperoxidase staining. (E) High-power view of MC nodule by scanning electron microscopy in cross section. (F) MC nodule count and collagen type I alpha1 (COL1a1), and collagen type IV alpha1 (COL4a1) RNA quantification (n=3). (G) Effect of ALK5 inhibition (Alk5i) on MC nodule formation (n=3). (H) Effect of SMAD2 or SMAD3 siRNA knockdown in MCs on nodule formation (n=3) with immunoblots demonstrating degree of knockdown. [DharmaFECT (DH), non-targeting siRNA control (siCP), siSMAD2 (siS2), siSMAD3 (siS3), total SMAD2 (tSMAD2), total SMAD3 (tSMAD3)]. Error bars represent 1 s.d. NS, not significant; ***, p<0.001; **, p<0.01: 2-tailed Student’s t test.

Article Snippet: After blocking, they were immunoblotted with rabbit anti-pSMAD2 (3108, Cell Signalling, MA, USA), rabbit anti-pSMAD1/5 (9516, Cell Signalling, MA, USA), rabbit anti-pSMAD3 (1880–1, Epitomics, Abcam, Cambridge, UK), rabbit anti-tSMAD2 (3122, Cell Signalling, MA, USA), rabbit anti-tSMAD3 (9513, Cell Signalling, MA, USA), or mouse anti-β actin (A5441, Sigma-Aldrich, MO, USA) for two hours (40 minutes for actin) at room temperature or overnight at 4°C.

Techniques: Staining, Immunoperoxidase Staining, Electron Microscopy, Inhibition, Western Blot