a Cell lysates from of U87-MG cells stably expressing shRNA against control or OGT and overexpressing HA-ACSS2-WT, HA-ACSS2-S267A, and HA-ACSS2-S267D were collected for immunoblot analysis with the indicated antibodies. b Representative images of cells generated in a and seeded into anchorage-independent growth assay and imaged at day 14. c Average colony formation quantified and presented as average from three independent experiments corresponding to b showing U87-MG cells stable expressing ACSS2-WT, ACSS2-S267A and ACSS2-S267D mutants and expressing shControl or shOGT as indicated. Student’s t-test reported as mean ± SD. *p-value < 0.05. d Representative images of tumor growth detected via bioluminescence (top) and H&E staining (bottom) U87-MG-luciferase cells subjected to the same treatment as in a and then used for orthotopic intracranial injections in mice. e Quantification of tumor size at Day 16 corresponding to d (shControl n = 5, shOGT n = 8, shOGT+ACSS2-S267A n = 4, shOGT+ACSS2-S267D n = 4). Student’s t-test reported as mean ± SD. *p-value < 0.05.