rabbit anti human integrin β1 polyclonal  (Cell Signaling Technology Inc)

Journal: Molecular medicine reports

Article Title: Sustained activation of C3aR in a human podocyte line impairs the morphological maturation of the cells.

doi: 10.3892/mmr.2020.11626

Figure Lengend Snippet: Figure 5. Overexpression of C3a significantly decreased the levels of VCL, PTK2 and ITGB1 in HPC cells. All cells (HPC, NC, C3a and SB) were cultured in the maturation condition for 6 days. Total proteins were extracted and (A‑C) western blotting and (D‑F) statistical analysis for VCL, PTK2 and ITGB1 was performed. *P<0.05, **P<0.01 vs. the HPC group; #P<0.05, ##P<0.01 vs. HPC‑NC group; &P<0.05, &&P<0.01 vs. the C3a group. C3a, human C3A anaphylatoxin; PTK2, protein tyrosine kinase 2; ITGB1, integrin β1; HPC, human podocyte cell line; NC, negative control; SB, SB290157; VCL, vinculin.

Article Snippet: Following blocking with 5% skim milk powder at room temperature for 4 h, the membrane was probed with primary antibodies, including rabbit anti-human c3ar polyclonal (cat. no. sc-20138; Santa cruz Biotechnology, inc.; dilution 1:1,000), rabbit anti-human integrin β1 polyclonal (iTGB1; cat. no. 9699S; cell Signaling Technology, inc.; dilution 1:1,500), rabbit anti-human vinculin (Vcl; cat. no. 26520-1-aP; ProteinTech Group, inc.; dilution 1:1,000), rabbit anti-human protein tyrosine kinase 2 (PTK2; cat. no. 71433; cell Signaling Technology, inc.; dilution 1:1,000) or mouse anti-human GaPdH monoclonal (cat. no. MB001; Bioworld Technology, Inc; dilution 1:10,000) antibodies at 4 ̊C overnight.

Techniques: Over Expression, Cell Culture, Western Blot, Negative Control

Journal: Molecular medicine reports

Article Title: Sustained activation of C3aR in a human podocyte line impairs the morphological maturation of the cells.

doi: 10.3892/mmr.2020.11626

Figure Lengend Snippet: Figure 6. Addition of C3a to the medium only partially mimics the effect of overexpression of C3a in HPC cells. HPC cells were divided into three groups: HPC, HPC+SB+C3a and HPC+C3a. Morphological changes were observed under a phase contrast microscope. (A) Reverse transcription‑PCR analysis of the expression of focal adhesion‑associated genes. *P<0.05, **P<0.01 vs HPC group; #P<0.05, ##P<0.01 vs HPC+C3a group. (B) Immunofluorescence staining for VCL. Scale bar, 50 µM. (C) Representative images taken under phase contrast microscopy demonstrated cell morphology. Scale bar, 100 µM. (D) Results of adhesion analysis. COL1A1, collagen I α1; FN1, fibronectin 1; LAMA1, laminin α1; LAMB1, laminin β1; LAMC1, laminin γ1; ITGA1, integrin α1; ITGA2, integrin α2; ITGB1, integrin β1; PTK2, protein tyrosine kinase 2; NRP1, neuropilin 1; TLN2, talin 2; FERMT2, fermitin family member 2; VCL, vinculin; C3a, human C3a anaphylatoxin; HPC, human podocyte cell; SB, SB290157.

Article Snippet: Following blocking with 5% skim milk powder at room temperature for 4 h, the membrane was probed with primary antibodies, including rabbit anti-human c3ar polyclonal (cat. no. sc-20138; Santa cruz Biotechnology, inc.; dilution 1:1,000), rabbit anti-human integrin β1 polyclonal (iTGB1; cat. no. 9699S; cell Signaling Technology, inc.; dilution 1:1,500), rabbit anti-human vinculin (Vcl; cat. no. 26520-1-aP; ProteinTech Group, inc.; dilution 1:1,000), rabbit anti-human protein tyrosine kinase 2 (PTK2; cat. no. 71433; cell Signaling Technology, inc.; dilution 1:1,000) or mouse anti-human GaPdH monoclonal (cat. no. MB001; Bioworld Technology, Inc; dilution 1:10,000) antibodies at 4 ̊C overnight.

Techniques: Over Expression, Microscopy, Expressing, Immunofluorescence, Staining