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. 2010 Jul;51(7):3599–3610. doi: 10.1167/iovs.09-4797

Figure 6.

Figure 6.

Effect of the IGFR inhibitor AG1204 on vitreous-induced lens fiber differentiation. (iA) Representative Western blots of explants cultured without (lanes 1 and 2) or with (10 ng/mL; lanes 3 and 4) IGF in the presence (lanes 2 and 4) or absence (lanes 1 and 3) of 5 μM AG1024 added 2 hours before growth factor treatment. AG1024 blocked IGF-induced ERK1/2 phosphorylation. (iB) Quantification of the relative density of phosphorylated ERK1/2 showed the same trend as the Western analysis. (iiA) Representative Western blots of explants cultured without (lanes 1 and 2) or with (lanes 3 and 4) FGF in the presence (lanes 2 and 4) or absence (lanes 1 and 3) of 5 μM AG1024 added 2 hours before growth factor treatment. AG1204 did not block FGF-induced ERK1/2 phosphorylation. (iiB) Quantification of the relative density of phosphorylated ERK1/2 that showed the same trend as the Western analysis. (iii) Representative micrographs of lens explants cultured without (A–D) or with (E–H) FGF, in the presence (B, D, F, H) or absence (A, C, E G) of 5 μM AG1024 added 2 hours before FGF treatment. AG1024 did not block FGF-induced lens fiber differentiation (F). (iv) Representative micrographs of lens explants cultured with vitreous and 5 μM AG1024. Explants were cultured without (A–D) or with (E–H) vitreous, in the presence (B, D, F, H, I, J) or absence (A, C, E, G) of 5 μM AG1024, added 2 hours before vitreous treatment. AG1024 reduced the accumulation of β-crystallin and cell elongation, but did not completely block the elongation. Some bare patches were observed over the explants (F, arrows), indicative of cell death/loss. Scale bar, (A–H) 50 μm; (I, J) 200 μm. (vA) Representative Western blots of explants cultured without or with vitreous in the presence or absence of 5 μM AG1024 added 2 hours before vitreous treatment, assayed for phosphorylated Akt (top), phosphorylated ERK1/2 (middle), and total ERK1/2 (bottom). Akt was phosphorylated within 20 minutes and maintained for up to 18 hours. The duration of ERK1/2 phosphorylation was reduced to 6 hours. Quantification of the relative density of phosphorylated Akt (vB) and ERK1/2 (vC) showed the same trend as the Western analysis.