Golgin Subfamily A Member 5 Is Essential for Production of Extracellular Matrix Proteins during TGF-β1-Induced Periodontal Ligament-Fibroblastic Differentiation

Figure 2

Identification of the antigenic molecule purified from the immunoprecipitates of anti-LG11 antibody. (a) The antigenic molecule recognized by anti-LG11 antibody. (A) Detection of the biotin-labeled LG11 antigenic molecule. Lane 1: preclearing without anti-LG11 antibody as control; lane 2: immunoprecipitates of anti-LG11 antibody. The antigenic molecule was indicated with arrowhead. (B) Identification of the antigenic molecule by mass spectrometric analysis. The tryptic peptides (underlines) were matched with GOLGA5 in protein sequence coverage of 46%. (b) Crossreactivity between anti-LG11 antibody and a commercial anti-GOLGA5 antibody in HeLa cells expressing GOLGA5 cDNA (A and B) and in TGF-β1-induced PDL fibroblastic cells (C). Full length cDNA construct tagged by FLAG was expressed in HeLa cells and detected by anti-FLAG (A), anti-LG11, and anti-GOLGA5 antibodies (B). Anti-FLAG immunoprecipitates (α-FLAG IP) recognized by both anti-LG11 (α-LG11 IB) and anti-GOLGA5 (α-GOLGA5 IB) antibodies. The input amount of total cell extract was indicated as HeLa CE. Lanes 1 and 3: vector only; lanes 2 and 4: cDNA transfection.

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