Fig. 6

CircRNF10 defends ferroptosis via upregulating HSPB1 in GSCs. a, b qPCR assays of the transcriptional levels of HSPB1 in circRNF10-overexpressed SYNS05 and SYNS06 (a) as well as circRNF10-knockdown SYNS03 and SYNS08 (b). c, d Western blotting showed the protein level change of HSPB1 after circRNF10 knockdown (c) or overexpression (d) in GSCs. e, f. GSH (e) and MDA (f) contents measured in SYNS03 and SYNS08 with circRNF10 knockdown followed by HSPB1 overexpression. g, h Lipid peroxidation levels (g) detected by BODIPY (581/591) C11 probe in circRNF10-knockdown SYNS03 and SYNS08 followed by HSPB1 upregulation and the relative fluorescence intensity of O-BODIPY quantified by Image J (h). Scale bar = 100 μm. i, Representative images of NSFA with circRNF10 knockdown in SYNS03 and SYNS08 followed by HSPB1 overexpression. Scale bar = 200 μm. j, k ELDA in SYNS03 (j) and SYNS08 (k) after circRNF10 silencing followed by HSPB1 overexpression. l Relative sizes of neurospheres of circRNF10-silenced SYNS03 and SYNS08 after HSPB1 upregulation. m, n Representative images of FerroOrange staining assays illustrated the intracellular Fe²⁺ accumulation of circRNF10-knockdown SYNS03 and SYNS08, followed by HSPB1 overexpression (m). The relative fluorescence intensity of FerroOrange probe quantified by Image J (n). Scale bar = 100 μm. o Representative images of cell death in SYNS03 and SYNS08 after 48 h of indicated treatments. Scale bar = 100 μm. p, q Real-time analysis of cell death in SYNS03 (p) and SYNS08 (q) with circRNF10 knockdown followed by HSPB1 upregulation. All data are expressed as the mean ± SD (three independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significance