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