Fig. 4

Gene Ontology (GO) analysis of the sunitinib gene signature in RB organoids. A, B The top 10 significant biological process terms enriched among differentially expressed genes (DEGs) of RB170 (A) and RB654 (B) organoids with sunitinib treatment at IC₁₀ (2 µM and 3 µM, respectively) for 72 h. Rich factor is defined as the ratio of input DEGs with a given annotated term to all genes with that annotation. C, D Gene networks of the top five enriched GO terms after treatment of RB170 (C) and RB654 (D) organoids with sunitinib. E, F Expression of JUN and retina-associated genes from a set of DEGs from RB170 (E) and RB654 (F) organoids. G, H Protein expression of c-Jun detected by western blotting in RB170 (G) and RB654 (H) organoids treated with vehicle or sunitinib at IC₁₀ and IC₅₀ (4 µM and 6 µM, respectively) for 72 h. Data obtained from three independent experiments; means ± SEM are shown. I, J Representative staining of c-Jun (red), RXRG (green), and nuclei (DAPI, blue) in RB170 (I) and RB654 (J) organoids treated with vehicle or sunitinib at IC₅₀. Image acquisition and processing was performed using the identical parameters. Scale bar: 50 μm (I, J)