Figure 4.

Largazole exerts anti-angiogenic effects through multiple signaling pathway. (A) 10 nM Largazole inhibits choroidal vessel outgrowth in cooperative manner with 500 µM Aflibercept. Representative images (left) and quantitative analysis (right) of microvessel formation from mouse choroidal explants (n = 3 independent experimental groups, n ≥ 6 explants per treatment group). Scale bar: 100 μm. All images shown are representative, and data are presented as means ± s.e.m. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test or unpaired, two-tailed Student’s t-test; * p < 0.05, and *** p < 0.001. (B) Largazole at concentration of 100 nM inhibits multiple angiogenic pathways. Representative western blots for VEGFR2, p21, pSMAD2/3, tSMAD2/3, and GAPDH in Largazole- and vehicle-treated HRECs (n = 3).