Figure 2
Inhibitory effects of an RAR-α agonist on the TGF-β2-induced expression of EMT markers in RPE cells. (a) RPE cells were cultured in collagen gels with or without TGF-β2 (1 ng/ml) and Am580 (10 µM) for 48 h, after which cell lysates were prepared and subjected to immunoblot analysis with antibodies to α-SMA and to β-tubulin (loading control). (b) The intensity of each α-SMA band in blots similar to that in (a) was normalized by that of the corresponding β-tubulin band, and the normalized values were expressed relative to that for control cells and are presented as means ± s.d from three independent experiments. The intensity of each immunoreactive bands was measured with the use of the Gels commands in ImageJ software. (c) Serum-deprived RPE cells were cultured in 24-well plates first with or without Am580 (10 μM) for 6 h and then in the additional absence or presence of TGF-β2 (1 ng/ml) for 48 h, after which the relative abundance of α-SMA, fibronectin, and collagen type I mRNAs was determined by RT-qPCR analysis. Data were normalized by the amount of GAPDH mRNA and are means ± s.d. from three independent experiments. **P < 0.01 (Dunnett’s test).
