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. 2024 Oct 24;27(3):695–710. doi: 10.1093/neuonc/noae224

Figure 3.

Figure 3.

ROR1 promotes GRB2/ERK/c-Fos signaling in GSCs. (A) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of differentially expressed genes between GSCs transduced with lentivirus expressing shNT or shROR1. (B) GSEA for MAPK pathway in ROR1-high compared to ROR1-low patients in CGGA database. NES, normalized enrichment score; FDR, false discovery rate. (C) Western blot analysis of ROR1, GRB2, KRAS, p-C-RAF, p-ERK, ERK, and c-Fos expression in T3359 and T456 GSCs transduced with lentivirus expressing shNT or shROR1. (D) Western blot analysis of ROR1, GRB2, KRAS, p-C-RAF, p-ERK, ERK, and c-Fos expression in T3359 and T456 GSCs with ROR1 overexpression. (E) GSEA for KEGG cell cycle pathway in ROR1-high comparing to ROR1-low (upper) and FOS-high comparing to FOS-low (bottom) patients in CGGA database. NES, normalized enrichment score; FDR, false discovery rate. (F) The protein levels of ROR1, c-Fos, p21, CCND1, CDK2, and CDK6 in T3359 and T456 GSCs transduced with lentivirus expressing shNT or shROR1. p21, CCND1, CDK2, and CDK6 are cell cycle related proteins. (G) Western blot analysis of ROR1, c-Fos, p21, CCND1, CDK2, and CDK6 expression in T3359 and T456 GSCs with ROR1 overexpression. p21, CCND1, CDK2, and CDK6 are key regulators of cell cycle. (H) Immunofluorescent co-staining of ROR1 (red) and c-Fos, CCND1, p21, CDK2, and CDK6 (green) in xenografts derived from T3691 GSCs transduced with lentivirus expressing shNT or shROR1. Scale Bar, 25 μm. The expression of c-Fos, CCND1, CDK2, and CDK6 decreased, whereas the p21 increased with ROR1 knockdown. (I) Statistical analysis of c-Fos, CCND1, p21, CDK2, and CDK6-positive staining in xenografts of (H). **P < .01; ***P < .001. Data are represented as mean ± SEM.