Figure 4

CTGF knock-down with shRNA inhibits serum-dependent cell proliferation of cardiac fibroblasts. Primary cardiac fibroblasts (P) were infected with lentiviruses encoding 2 different CTGF shRNAs (h2 and h5 from OpenBiosystems) or with a lentivirus generated from an empty vector (V) as control. Panel A: CTGF protein expression was measured by ELISA in culture supernatants. Panel B: 96 hrs after infection, cells were re-plated in quadruplicate wells of a 96-well plate in medium containing serum and defined growth supplements and incubated for up to 10 days. Cell proliferation was measured by Cyquant Fluorescense Intensity at the indicated timepoints. Panel C: Parallel cultures of cells were stained with crystal violet at day 10 and doubling time (DT) was determined by linear regression analysis on the data shown in B. * p < 0.001 vs. vector by ANOVA.