Fig. 3. Collagens-integrins signaling is essential for AR to enhance the migration potential of PDCs in vitro.
Primary PDCs were isolated from ARflox/Y and AR-/Y;Prrx1::Cre mice. The AR in PDCs from ARflox/Y mice was knocked down using AR-targeted siRNA (si-AR). D1 cells were transfected with Flag (pcDNA3-flag) or Flag-tagged AR (pcDNA3-flag-AR). A, B Itgb1 and Itga2 mRNA levels were quantified by RT-qPCR. C, D Cells were cultured with or without the integrin ligand, Col I. E, F Cells were incubated with or without the integrin α2β1 inhibitor TC-I or integrin α2β1 blocking antibody CD49b. (C–F) Migration capacity was measured using the OrisTM Cell Migration Assay (CMA) kit. G D1 cells were transfected with pSG5 or AR (pSG5-AR). The integrin β1 in the transfected cells was knocked down using Itgb1-targeted siRNA (si-Itgb1). Cell migration assay was stained with Giemsa stain. AR, integrin β1 and GAPDH protein levels were then determined by western blot analysis. Experiments were conducted three times. Data were expressed as the mean ± SD. Statistical correlation of data was checked for significance by Student’s t test. *P < 0.05, **P < 0.001 was considered to indicate a statistically significance result.