Fig. 6: Erianin promotes TMZ sensitivity in TMZ-resistant GSCs through REST.

a–d Western blotting and qPCR assays showing REST protein (a, b) and mRNA expression (c, d) in normal, TMZ resistant, and REST overexpressed U87R and GSCm01R. e, f Cell viability measured with CCK-8 in REST overexpressed U87R and GSCm01R after combined treatment with TMZ and erianin. g, h Representative EdU assay showing proliferation and quantitative analysis in REST overexpressed U87R and GSCm01R after combined treatment with erianin and TMZ. Scale bar = 50 μm. i, k Migration assays of REST overexpressed U87R and GSCm01R using a Transwell 24-well plate without extracellular matrix gel after combined treatment. Scale bar = 50 μm. j, l Invasion assays of REST overexpressed U87R and GSCm01R using a Transwell 24-well plate with extracellular matrix gel after combined treatment. Scale bar = 50 μm. m, n ferroptosis detection by BODIPY (581/591) C11 probe in U87R and GSCm01R with combined treatment after REST overexpression and quantification of relative fluorescence intensity by Image J. Scale bar = 100 μm. o, p NSFA revealing the size of neurospheres and quantitative analysis of REST overexpressed GSCm01 after combined treatment with TMZ and erianin. Scale bar = 50 μm. q ELDA showing the neurosphere-forming ability of REST overexpressed GSCm01R after combined treatment with TMZ and erianin. r Western blotting of stemness marker in REST overexpressed GSCm01R after combined treatment with TMZ and erianin. All data are shown as the mean ± SD (five independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001; ns no significance.