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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Correction: CircLRFN5 inhibits the progression of glioblastoma via PRRX2/GCH1 mediated ferroptosis

Fig. 4

CircLRFN5 binds to PRRX2 protein and promotes its degradation via the ubiquitin-mediated proteasomal pathway. a CircLRFN5 binds to PRRX2 proteins via CatRAPID prediction. b-d RIP assays showed anti-PRRX2 proteins could lead to the enrichment of circLRFN5 in circLRFN5 overexpressed (b) or knockdown (c,d) GSCs. e RNA pull-down assays showed the biotinylated circLRFN5 probe pulled down PRRX2 proteins in GSC51 (left) and GSC53 (right). f Five biotinylated probes (1- 590, 590- 1085, 653- 1085, 1085- 1581 and 1148- 1581) containing different fragments of circLRFN5 were designed for RNA pull-down assay. g qPCR assays showed the mRNA expression of PRRX2 after circLRFN5 overexpression (left) or knockdown (right) in GSCs. h Western blotting showed the protein expression of PRRX2 after circLRFN5 overexpression (left) or knockdown (right) in GSCs. i, j The circLRFN5 overexpressed GSC51 (i) or knockdown GSC53 (j) were treated with cycloheximide (CHX, 100 ng/ml) and the half-life of PRRX2 protein was detected by western blotting. k The circLRFN5 overexpressed GSC51 (upper) or knockdown GSC53 (lower) was treated with or without MG-132 (50 μM) for 6 h, and PRRX2 expression was detected by western blotting. l, m In vitro ubiquitination assays showed the level of ubiquitination of PRRX2 protein after circLRFN5 overexpression (l) or knockdown (m) in GSCs. All data are expressed as the mean ±SD (three independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001

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