Fig. 2

Circ-hnRNPU inhibits O- and N-glycosylation in gastric cancer cells. a, Volcano plots (left panel), Venn diagram (middle panel), heatmap (middle panel), and GSEA (right panel) showing the altered gene expression and corresponding biological processes in MKN-45 and AGS cells stably transfected with empty vector (mock) or circ-hnRNPU. b, Regulatory network (left panel) and ChIP-X analysis (right panel) showing the potential transcription factors for altered glycosyltransferase genes. c, Dual-luciferase assay revealing the activity of potential transcription factors in AGS cells stably transfected with mock or circ-hnRNPU (n = 5). d, ChIP and qPCR (upper panel) and ChIP-seq peak (GSE51334, lower panel) indicating endogenous enrichment of c-Myc on promoter regions of glycosyltransferases in AGS and HeLa cells. e, ChIP and qPCR (left panel) and real-time qRT-PCR (right panel, normalized to β-actin) assay showing the c-Myc enrichment and transcriptional levels of GALNT6 and MGAT1 in AGS and HeLa cells stably transfected with mock or circ-hnRNPU (n = 4). f, Western blot assay indicating the levels of O- and N-glycosylation in AGS and NCI-N87 cells stably transfected with mock, circ-hnRNPU, scramble shRNA (sh-Scb), or sh-circ-hnRNPU. Student’s t test compared the difference in c-e. *P < 0.05, **P < 0.01. Data are shown as mean ± s.e.m. (error bars) or representative of three independent experiments in c-f