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Figure 5


Fig. 5. Effects of MEK-ERK inhibition on VEGF-dependent proliferation and differentiation. (A) Western blots of VEGF-induced phospho-ERK. E6 retinas were cultured with or without U0126 for 60 minutes before VEGF was added for 10 minutes. Blots were probed with phospho-ERK1/2 (top) and {alpha}-tubulin (bottom) antibodies. (B) Quantification of optical densities of phospho-ERK signals were normalized against {alpha}-tubulin signals (n=4). (C) RT-PCR detection of cyclin D1 transcripts in E6 retinas cultured for 24 hours in the presence or absence of VEGF. Ratios of cyclin D1 and GAPDH products are shown (n=8). (D-F) Quantifications of U0126 effects on VEGF-dependent cell proliferation (D) or differentiation (E,F). E5 explants were cultured for 24 hours and BrdU labeled for the last 3 hours (n=6; *P<0.03).