Fig. 1. Effects of Ell3-suppressed ADSC-CM on the proliferation, migration and migration-related marker expression of mouse embryonic fibroblasts (MEF) in vitro.

(A) siRNA-mediated suppression of Ell3 in ADSCs. Ell3 expression was quantitatively analyzed by real-time RT-PCR 48 hours after siRNA transfection. (B) The proliferation of MEF cultured in CM collected from siNS- or siEll3-transfected ADSCs was analyzed by counting cells on the indicated days after seeding. (C) The migration ability of MEF cultured in CM collected from siNS- or siEll3-transfected ADSCs was analyzed by the scratch/wound assay. Cell migration was photographed on the indicated days after scratching (left), and the relative migration ability was quantified by subtracting the cell-free areas on days 1 and 2 from the cell-free area on day 0 (right). (D) The migration ability of MEF was analyzed by the transwell assay as described in the Materials and Methods. Images were captured at 24 hours under 40× magnification (left). Cells were counted from 10 randomly selected fields, and the averages were calculated (right). (E) Relative expression of MMP family genes in MEF treated with CM collected from ADSCs transfected with siNS or siEll3 for 24 hrs. Abbreviations: siNS, non-specific siRNA; siEll3, siRNA targeting Ell3; CM, conditioned media; SFM, serum-free media; MEF, mouse embryonic stem cells. The error bars represent the standard errors from three independent experiments, which were each performed using triplicate samples. * P<0.05 (Student’s t-test).