boster rabbit anti α sma  (Boster Bio)

Journal: PLoS ONE

Article Title: TRPV4 Channel Inhibits TGF-β1-Induced Proliferation of Hepatic Stellate Cells

doi: 10.1371/journal.pone.0101179

Figure Lengend Snippet: A. The level of the TRPV4 was analyzed by immunohistochemistry in human normal liver and liver fibrosis patients. Representative views from each group are presented (original magnification, ×50). B. Liver tissues from liver fibrosis patients tissue were double stained with TRPV4 and α-SMA antibodies. Representative photomicrographs are shown (original magnification, ×40) in B. C. Pathology observation of the experimental rat liver sections stained with hematoxylin and eosin (H&E) staining and massion staining (×200). D. The level of the α-SMA was analyzed by immunohistochemistry in vehicle control livers and liver fibrotic tissue. Representative views from each group are presented (original magnification, ×50). E-F. Total RNAs were isolated from the livers of CCl 4 -treated rats or vehicle-treated groups. The expression of TRPV4 mRNA was assessed by RT-PCR(E) and realtime PCR(F). Representative images of three independent experiments are shown. *p<0.05, **p<0.01 vs. vehicle control. G. Whole-cell extracts were isolated from the liver tissues of CCl 4 -treated rats or vehicle-treated groups, and subjected to immunoblot for TRPV4 and β-actin control. Representative images of three independent experiments are shown. *p<0.05, **p<0.01 vs. vehicle control.

Article Snippet: Anti-TRPV4 (Cell Signaling, Beverly, MA, USA) was diluted 1∶400, and rabbit monoclonal anti-α-SMA (Boster) was diluted 1∶200.

Techniques: Immunohistochemistry, Staining, Isolation, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: PLoS ONE

Article Title: TRPV4 Channel Inhibits TGF-β1-Induced Proliferation of Hepatic Stellate Cells

doi: 10.1371/journal.pone.0101179

Figure Lengend Snippet: A. Total RNAs were isolated from TGF-β1-treated HSC-T6 cells, and subjected to qRT-PCR analyses. Representative images of three independent experiments are shown. *p<0.05 vs. non-treated cells. B. Whole-cell extracts were isolated from TGF-β1-treated HSC-T6 cells, and subjected to Western blot analyses with TRPV4 and β-actin antibodies. Representative blots of three independent experiments are shown. **p<0.01 vs. non-treated cells. C. Total RNAs were isolated from TGF-β1 treated HSC-T6 cells at different time points. The expression of α-SMA and Col1a1 mRNA was assessed by RT-PCR. Representative images of three independent experiments are shown. **p<0.01 vs. non-treated cells.

Article Snippet: Anti-TRPV4 (Cell Signaling, Beverly, MA, USA) was diluted 1∶400, and rabbit monoclonal anti-α-SMA (Boster) was diluted 1∶200.

Techniques: Isolation, Quantitative RT-PCR, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: TRPV4 Channel Inhibits TGF-β1-Induced Proliferation of Hepatic Stellate Cells

doi: 10.1371/journal.pone.0101179

Figure Lengend Snippet: A. Total RNA extracts were made from HSC-T6 cells treated with or without TGF-β1 and Ru, and subjected to qRT-PCR analyses of TRPV4. Representative images of three independent experiments are shown. # p<0.05 vs. TGF-β1-treated cells. B. HSC-T6 cells were seeded in triplicate on day 0 and incubated in DMEM containing 10% fetal bovine serum or same media supplemented with Ru for further 24 h. Proliferation was measured by adding 5 mg/ml MTT reagent per well and incubating it for 4 h. # p<0.05 vs. TGF-β1-treated cells. C. Total RNA extracts were made from HSC-T6 cells treated with or without TGF-β1 and Ru, and subjected to qRT-PCR analyses of α-SMA. Representative images of three independent experiments are shown. # p<0.05 vs. TGF-β1-treated cells. D. Whole-cell protein extracts were made from HSC-T6 cells treated with or without TGF-β1 and Ru, and subjected to Western blot analyses of TRPV4. Representative images of three independent experiments are shown. ## p<0.01 vs. TGF-β1-treated cells. E. HSC-T6 cells were treated with TGF-β1 for 48 h, followed by transfection with TRPV4-siRNA for an additional 48 h, and cell viability was determined by MTT assay. Mean±SE of two HSC preparations in quadruplets is shown; *p<0.05 vs. non-treated cells, # p<0.05 vs. TGF-β1-treated cells. F. Whole cell extracts were isolated from TGF-β1-treated HSC-T6 cells with RNAi transfection, and subjected to Western blot analyses. Representative images of three independent experiments are shown. **p<0.01 vs. non-treated cells, ## p<0.01 vs. TGF-β1-treated cells.

Article Snippet: Anti-TRPV4 (Cell Signaling, Beverly, MA, USA) was diluted 1∶400, and rabbit monoclonal anti-α-SMA (Boster) was diluted 1∶200.

Techniques: Quantitative RT-PCR, Incubation, Western Blot, Transfection, MTT Assay, Isolation