Fig. 4

Impairment of Igf2bp1 increases cell death and layering defects in the retina.

Tg(isl2b:GFP)^zc7 stable transgenic embryos (a-h) were uninjected (a, e) injected with p53 MO (b, f), e3i3 (c, g), or co-injected with p53 MO+e3i3 MO (d, h). Embryos were fixed and stained with α-EGFP (green) and TO-PRO-3 (red in a-d, gray in e-h). Images are maximum intensity projections (a-d, i-l) or singles slices (e-h) of lateral views of eyes, with lens removed, taken with a confocal microscope (20x lens). Morphant eyes were missing RGCs in central retina (white arrowheads in c, d) and had displaced RGCs (yellow arrowheads in c, d). (g, h) Single z-slices have holes (white arrowheads) and abnormal layers (yellow arrowheads). (i, l) Maximum intensity projections, lateral view of AO staining in 31 hpf embryos, within the lens (inner dotted yellow circle) and retina (outer dotted yellow circle). (m) The AO-positive cells were counted in retinas alone (not including lens and extraocular tissues) from; uninjected (n = 10), p53 MO injected (n = 9), e3i3 MO injected (n = 10), and coinjected with p53 MO and e3i3 MO (n = 10) embryos. A one-way ANOVA (p<0.0001) with Tukey HSD test (p<0.01) showed that e3i3 MO injected embryos and embryos coinjected with e3i3 MO and p53 MO were both significantly different than controls, but not significantly different from each other. The black points on the graph represent mean +/- SEM. Scale bars are 50 µm.