Fig. 5

ZBTB48 transcriptionally upregulates HSPB1 expression in GSCs. a Correlation analysis of HSPB1 and ZBTB48 based on TCGA and CGGA datasets. b Differentially expressed genes obtained from TCGA and CGGA datasets divided into the low expression group and high expression group based on the median expression of ZBTB48 and GSEA plots depicting indicated ferroptosis signaling pathway in low expression group. c-f, qPCR assays of the transcriptional levels of HSPB1 in ZBTB48-silenced SYNS03 and SYNS08(c) as well as ZBTB48-overexpressed SYNS05 and SYNS06 (d). Western blotting showed the protein level change of HSPB1 after ZBTB48 knockdown (e) or overexpression (f) in GSCs. g Left schematic represents ZBTB48 binding motif obtained from AnimalTFDB database. Right schematic diagram shows the two putative ZBTB48 binding sites and matched mutant sequences in the HSPB1 promoter region for Dual-luciferase reporter assays. h–k The Dual-luciferase reporter assays revealed the luciferase promoter activities of HSPB1 with ZBTB48 scliencing in SYNS03 (h) and SYNS08 (i) or ZBTB48 overexpression in SYNS05 (j) and SYNS06 (k). l, m The ChIP qPCR showed that anti-ZBTB48 treatment could detect the enrichment difference of HSPB1 promoter sequence in ZBTB48-silenced SYNS03 and SYNS08 (l) or in ZBTB48-overexpressed SYNS05 and SYNS06 (m). All data are expressed as the mean ± SD (three independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significance