Fig. 2

Propofol increases EZH2 palmitoylation mediated by ZDHHC5. A RT-PCR analysis of the mRNA levels of 24 known DHHCs in U118-derived GSCs treated with or without propofol (n = 5 in each group). B Lysates of propofol-treated U118-derived GSCs were subjected to the acyl-biotinyl exchange method (ABE) with a streptavidin biotinylated antibody. Palmitoylated proteins were subjected to LC–MS/MS analysis. The identified peptide sequences of EZH2 are shown. C Acyl-biotinyl exchange method and western blotting for palmitoylated EZH2 and western blotting for ZDHHC5, p-EZH2 S21, and H3K27me3 in U118-derived GSCs treated with or without propofol (n = 5 in each group). GAPDH was used as a loading control. D The mRNA levels of CDKN1B, RUNX3, and HOXA5 in U118-derived GSCs treated with or without propofol were analyzed by RT-PCR (n = 5 in each group). β-actin was used as the loading control. E Western blotting for CDKN1B, RUNX3, and HOXA5in U118-derived GSCs treated with or without propofol (n = 5 in each group). GAPDH was used as a loading control. F Detection of glioma spheres formed for U118-derived GSCs treated with or without propofol, and EZH2 inhibitor, tazemetostat, and cultured for a second passage. Scale bars, 100 μm. Quantification of glioma spheres formation capacity (n = 5 in each group). Ctrl, control; Taz, tazemetostat