Fig. 1

Overexpression of GD2 in NB tumor tissues is mediated by B3GALT4. A Immunohistochemistry (IHC) analysis of GD2 expression in ganglioneuroma (GN) (n = 25) and neuroblastoma (NB) tumor tissues (n = 81). Scale bar: 100 μm. B The correlation between the International Neuroblastoma Staging System (INSS) stage and GD2 expression of NB was analyzed. C IHC scores were applied to assess GD2 expression in NB specimens with or without bone marrow metastasis. D The correlation between the Children’s Oncology Group (COG) stage and GD2 expression of NB was analyzed. E Kaplan-Meier survival curve for 3-year overall survival of 60 NB patients in the 81 patients (Fig. 1A-D) according to GD2 expression levels (log-rank test and the p-value is shown). F B3GALT4 was a crucial enzyme for ganglioside GD2 synthesis. G Gene set enrichment analysis (GSEA) analysis showed that B3GALT4 overexpression was enriched in hydrolyzing N-glycosyl compounds based on data from the GEO database (GSE49710). H Differences in B3GALT4 level between GD2 (+) and GD2 (−) NB samples from the GEO databases (GSE90689 and GSE112447). I Negative correlation between the expression level of GD2 and B3GALT4 in NB tissues (n = 25). Scale bar: 100 μm. J, K B3GALT4 knockdown and overexpression 9464D and 975A2 stable cell lines were constructed using recombinant lentiviral transfection. The overexpression and knockdown efficiency were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. Quantitative data are shown as means ± SEM from three independent experiments. Representative Western blots and bar charts are shown. (L) Representative flow cytometric analysis images of GD2 levels in 9464D and 975A2 cells with different treatments and statistical analysis results