Extended Data Fig. 7: Activation of HIF-1α in RPE induces type-I CNV-like phenotype in 3D-oBRB.
a-h, immunostaining of tissues with anti-CD31 (magenta), anti-HIF-1α (yellow) antibodies, and staining for F-ACTIN (cyan), and nuclei (blue). Tissues were treated with either vehicle (DMSO, a,e), or ML228 (2 μM) on apical side of RPE only (b,f), or basal side of choroid only (c,g), or both sides (d,h). Red arrowheads indicate type-I CNV-like phenotype, white arrowheads indicate examples of HIF-1α translocation to the nuclei. (n = 3), scale bars, 50 μm. i, TER measurements normalized to ctrl. (n = 6). Data are presented as mean values +/− SEM. ANOVA and Tukey’s multiple comparisons test. j, k, pseudo 3D-projected side views of 3D-oBRB immunostained with anti-CD31 (magenta), anti-COL IV (yellow), anti-LAMININ (cyan) antibodies, and nuclei (blue). Tissues were treated with either vehicle (DMSO, j), or ML228 (2 μM) on apical side of RPE only (k). Arrowheads indicate type-I CNV-like phenotype in ML228-treated (k) tissues with choriocapillaris (CC) penetrating through the Bruch’s membrane (BM). (n = 3), scale bars, 10 μm.
