Fig. 6

CircLRFN5 downregulates PRRX2 mediated transcription of GCH1 in GSCs. a A Venn Diagram showed the correlation between PRRX2 and ferroptosis-related genes in TCGA and CGGA datasets (R more than 0.4). b, c The correlation between PRRX2 and GCH1 in TCGA (b) and CGGA (c) datasets. d Sequence motif representing the consensus PRRX2 binding motif according to the JASPAR database. e Schematic diagram of the putative PRRX2 binding site in the promoter of GCH1. f–h Western blotting (f) and qPCR (g, h) showed the expression of GCH1 in circLRFN5 overexpressed GSC51 and GSC58, followed by PRRX2 overexpression. i, j The luciferase reporter assays showed the luciferase promoter activities of GCH1 after circLRFN5 overexpression in GSC51 (i) and GSC58 (j), followed by PRRX2 overexpression. k, l The ChIP qPCR showed that anti-PRRX2 treatment could lead to the enrichment of GCH1 in circLRFN5 overexpressed GSC51 (k) and GSC58 (l), followed by PRRX2 overexpression. All data are expressed as the mean ± SD (three independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001