Fig. 3

CircLRFN5 inhibits GSCs viability and proliferation in a ferroptosis-dependent manner in vitro. a Cell viability assays showed whether VAD (20 μM), Nec (20 μM), 3-MA (60 μM) and Fer-1 (10 μM) could rescue the growth inhibition of GSC51 induced by circLRFN5 overexpression after 48 h. b, c Representative images of BODIPY (581/591) C11 staining in different groups of GSCs and the relative fluorescence intensity of O-BODIPY was quantified by image J. Scale bar = 50 μm. d, e GSH (d) and MDA (e) levels were detected in circLRFN5 overexpressed GSC51 and GSC58, followed by Fer-1 treatment. f, g MTS assays showed the cell viabilities of circLRFN5 overexpressed GSC51 (f) and GSC58 (g), followed by Fer-1 treatment. h, i Representive EdU assays showed the proliferation of circLRFN5 overexpressed GSC51 and GSC58, followed by Fer-1 treatment. Scale bar = 50 μm. j Representive neurospheres formation assays showed the relative sizes of neurospheres of circLRFN5 overexpressed GSC51 and GSC58, followed by Fer-1 treatment. Scale bar = 20 μm. k ELDA assays showed the neurospheres formation abilities of circLRFN5 overexpressed GSC51 (left) and GSC58 (right), followed by Fer-1 treatment. l Western blotting detects the stemness markers of circLRFN5 overexpressed GSC51 and GSC58, followed by Fer-1 treatment. m Cell cycle assays showed the cell cycle distributions of circLRFN5 overexpressed GSC51 and GSC58, followed by Fer-1 treatment. All data are expressed as the mean ± SD (three independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001