Fig. 2: AGGF1 is elevated in the retinas of the OIR mice and co-localized with neovascular endothelial cells.

a Schematic illustration of the mouse oxygen-induced retinopathy (OIR) model. b FITC-dextran staining of whole-mount retinas from normoxic and OIR mice at P12, P17, and P21 (n = 3 mice per group). In the inserts, the red line represents the edge of the retina, the blue area represents the avascular area, and the red area represents neovascular tufts (NVTs). c, d Western blot analysis and quantification of AGGF1 and VEGF protein levels in the OIR retinas at P12, P17, P21 (n = 4 mice per group). e Immunofluorescence staining for AGGF1 on retinal sections at P12, P17, and P21 of OIR mice (n = 4 mice per group). AGGF1 is expressed by blood vessels in the superficial, intermediate, and deep layers. f Schematic illustration of the structure of retinal layers and the distribution of vessels in retinal sections. g Double immunofluorescence staining for AGGF1 (red) and CD31 (green) in P17 OIR retinas (n = 4 mice per group). h Localization and quantification of AGGF1 protein in whole-mount retinas upon normoxia and OIR at P17 (n = 5 mice per group). i Schematic illustration of the vascular network in flat-mounted retinas. j Immunofluorescence for AGGF1 and isolectin B4 (IB4) in whole-mounted retinas of OIR mice at P17 (n = 5 mice per group). AGGF1 co-localized with IB4 in microvessels and large blood vessels. k Immunofluorescence for AGGF1 and Iba-1 (a microglial marker) and GFAP (an astroglial marker) in whole-mount retinas of OIR mice at P17 (n = 5 mice per group). Error bars represent mean ± SEM. One-way ANOVA with Tukey’s multiple comparisons test (d). Scale bars: 1000 μm (b), 50 μm (e, g) and 100 μm (h, j, k); magnified images: 50 μm (g). GCL ganglion cell layer, INL inner nuclear layer, ONL outer nuclear layer. Source data are provided as a Source Data file.