Figure 3

Figure 3

Mouse RPE-choroidal fragments were cultured in Matrigel^® for 24 h and further cultured with 10 μM 22-oxacalcitriol for 6 days. On day 7, samples were fixed and sprouting area quantified from phase contrast images using ImageJ freehand tool. (A) 22-oxacalcitriol and positive control sunitinib significantly attenuated choroidal sprouting angiogenesis. Graph showing mean percent sprouting area compared to vehicle control ± SEM; statistical analyses were performed using one-way ANOVA with Dunnett’s post hoc test; asterisk indicates ^*p≤0.05 and ^***p≤0.001 and n = 3 with up to 6 replicates per individual experiment. (B) Representative brightfield images of mouse choroidal sprouting after 7 days with indicated treatments. (C) Representative calcein stained images of mouse choroidal sprouting after 7 days with indicated treatments. Calcein staining of RPE-choroidal cultures confirms explant viability in vehicle and 22-oxacalcitriol treated explants. Scale bar represents 0.5 mm.