Fig 1: A2E induces an increase of bFGF mRNA at transcriptional level in polarized ARPE19 cells. ARPE19 were cultured as described in material and methods before treatment with 5 μM A2E for 48 h. (A) Quantitative PCR shows the different mRNA levels of bFGF in ARPE19 cells treated or not with 5 μM A2E or 5 μM U0126. Quantification of the mRNA bFGF/GAPDH ratio is expressed as mean ± SEM of 3 experiments (n = 7–14) with 100% fixed for the untreated cells; **p < 0.007, ***p < 0.0001. (B) Western blot shows the expression level of bFGF (nuclear and secreted forms). Positive control was done using recombinant bFGF [SIGMA, F0291]. Quantification of the bFGF/TUBULIN ratio was expressed as mean ± SEM of 3 experiments (n = 9) with 100% fixed for the untreated cells; *p < 0.01. (C) Immunostaining of bFGF protein in non-permeabilized polarized ARPE19 cells after 48 h of exposure to 5 μM A2E, result is representative of two distinct experiments. (D) Western blot shows the expression level of type 1 COLLAGEN normalized by TUBULIN. Quantification of the COLLAGEN1/TUBULIN ratio was expressed as mean ± SEM of 3 experiments (n = 9) with 100% fixed for the untreated cells.