Fig. 3: Remodeling of glycosylation by lead compounds is independent of the glucocorticoid receptor (GR).

a Chemical structure of NSC80997 and NSC255112. b Representative images of MCF7^WT cells treated with DMSO, 10 µM or 10 nm NSC80997 or dexamethasone for 24 h, stained with isotype or anti-GR antibody (green) and DAPI (magenta). Scale bar represents 20 µm. c Bar diagram shows cell surface VVA binding to HEK293^WT cells treated for 24 h with increasing concentrations of mifepristone, mometasone furoate, cortisone, cortisol, dexamethasone, or NSC80997. DMSO and methanol served as controls. Data was assessed by flow cytometry, is presented as mean fluorescence intensity (MFI) (arbitrary units) and is representative of two independent experiments. d NSC80997-mediated Tn induction in GR knock-out (Δ) cell lines. Bar diagram shows VVA binding to HEK293^WT, HEK293^{KO NR3C1} and HEK293^{KO NR3C4} cells treated with 10 µM NSC80997 or DMSO control for 24 h. Lectin binding was assessed by flow cytometry and data are presented as average MFI values (arbitrary units) of two independent experiments. e The methyl ketone in NSC80997 was reduced to a secondary alcohol in NSC80997-KR highlighted in red. f Bar diagram shows VVA binding to HEK293^WT cells treated with increasing concentrations of NSC80997 or the ketone reduced derivative (NSC80997-KR). VVA binding was measured by flow cytometry and is presented as average MFI ± SEM from three independent experiments. ***P < 0.0001, **P < 0,005 (t-test).