Fig. 7

IGF2BP3 maintain the stability of circRNF10 and is transcriptionally upregulated by ZBTB48 in GSCs. a IGF2BP3 was predicted by RBP suite to interact with circRNF10. b, c RNA pull-down assays showing the biotinylated circRNF10 probes pull-down IGF2BP3 protein in SYNS03 (b) and SYNS05 (c). d, e RIP assays showing anti-IGF2BP3 treatment enriched with circRNF10 after circRNF10 overexpression (d) or knockdown (e). f, g qPCR showing circRNF10 expression after knockdown (f) or overexpression (g) of IGF2BP3 in GSCs. h, i RNA stability assays showing the half-life of circDNF10 in IGF2BP3-silenced (h) or overexpressed (i) GSCs followed by actinomycin D treatment. j Left schematic represents ZBTB48 binding motif and promoter region of IGF2BP3. Right schematic diagram shows the two putative binding sites and matched mutant sequences in the promoter region for Dual-luciferase reporter assays. k, l The Dual-luciferase reporter assays revealed the luciferase promoter activities of IGF2BP3 with ZBTB48 silencing in SYNS03 (k) and ZBTB48 overexpression in SYNS05 (l). m The ChIP qPCR showing the enrichment difference of IGF2BP3 promoter sequence via anti-ZBTB48 treatment in ZBTB48-silenced SYNS03 and SYNS08. n-q qPCR and Western blotting assays displayed the expression level’ s change of IGF2BP3 in ZBTB48-silenced (n, o) and ZBTB48-overexpressed GSCs (p, q). All data are expressed as the mean ± SD (three independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significance