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

Figure 1

Antigenic molecule recognized by a novel anti-LG11 antibody was upregulated in the TGF-β1-induced fibroblastic cells. (a) Cell morphological characterization. The morphologies (A) and cytoskeletal structures (B) of hPDLSC were observed before and after TGF-β1 treatment. The cytoskeletal structure was shown as F-actin by phalloidin staining. (b) Evidence of successful PDL-fibroblastic differentiation derived from hPDLSCs. PDL-fibroblastic and osteo/cementoblastic marker expressions in hPDLSCs were verified by transcriptional levels. Quantitative RT-PCR analysis is performed to determine the gene expression in cells treated with TGF-β1 (TGF) and/or its receptor inhibitor, SB431542 (SB). Statistical significance is determined using the student’s -test. ns: not significant; ; . (c) Antigen recognized by anti-LG11 antibody as a PDL fibroblast-specific molecule. (A) LG11 antigen specifically interacted with TGF-β1-induced PDL-fibroblastic progenitors (TGF), not with SB431542-treated cells (SB). (B) TGF-β1-induced PDL-fibroblastic progenitors were double positive of Cy3-PLAP-1 and FITC-LG11. 1: undifferentiated cells; 2: SB431542-treated hPDLSCs; 3: TGF-β1-induced PDL-fibroblastic progenitors. In (C), the amount of the endogenous LG11 antigen was significantly increased in total cell extract obtained from PDL-fibroblastic progenitors. (d) Sequence information and identity analysis of the variable regions of anti-LG11 antibody. (A) Amino acid sequences of mouse immunoglobulin heavy and light chain variable regions. Three complementarity determining regions in heavy and light chains (CDR-H1~3 and CDR-L1~3) were shown in shading boxes. (B) IMGT/V-QUEST database-based antibody similarity analysis.

(a)

(b)

(c)

(d)