Fig. 8: ALK5 inhibitor attenuates EC proliferation and the retarded vessel growth in Tfam^ECKO retinas.

a A diagram for the ALK5 inhibitor protocols. WT and the mutant pups were received i.p. injection of vehicle or ALK5 inhibitor LY364947 compound at 5 μg/g body weight daily from P2-P6 (Protocol 1) or P2-P14 (Protocol 2). Mice were subjected to analyses at P7 or P15. b, c Retina lysates were subjected to Western blotting with respective antibodies. Protein levels were quantified and presented as fold changes by vehicle WT as 1.0. Ratios of p-SMAD2:SMAD2 are presented as a graph (c). Data presented from one of 3 mice per group. d, e P7 retinas were subjected to whole mount staining for CD31. Vascular progression (the distance of radial extension of the vascular plexus from the optic center) was quantified (e). n = 6 mice per group. f, h Vehicle or LY364947 treated WT and Tfam^ECKO mice at P7 were injected with EdU (100 μg per gram of body weight). 4 h after i.p. injection of EdU, retinas were harvested and subjected to whole-mount staining with ERG (red) and CD31 (blue) followed by the EdU Click-iT assay (green). Arrows and arrowheads indicate EdU⁺ and EdU^- EC, respectively. EC density (ERG⁺/mm² retinal area) and EC proliferation (EdU⁺ERG⁺/total ERG⁺ ECs) were quantified by taking WT vehicle as 1.0 (g, h). n = 6 mice per group. Data are means ± SEM. P values are indicated, using one-way ANOVA followed by Tukey’s multiple comparisons test. Scale bar: 100 μm (b); 25 μm (d, g). Source data are provided as a Source Data file.