Fig. 2

Pharmacological inhibition of OGT arrests osteoclastogenesis at early stages of differentiation. Western blot analysis (a), representative images of high-content imaging, and cellomics analysis (b) of O-GlcNAcylation during in vitro osteoclastogenesis in the presence of TNFα and with or without OSMI-1 (n = 5 per group for western blot; n = 11 759 cells in total for cellomics). c Levels of osteoclast-related genes analyzed by real-time PCR after OSMI-1 treatment (n = 4 per group). d Representative images of high-content imaging and the cellomics analysis of the proportions of immature and mature osteoclasts, the OC fusion index, and TRAP enzyme activity (n = 102 666 cells in total). e Proliferation assay (n = 4 per group). f In vitro bone resorption assay with OSMI-1 treatment (n = 6 per group). All bar graphs are shown as median ± IQR. Horizontal scale bars are represented as 50 µm in IF images, 100 µm in bone resorption assay. Statistical significance was determined by Mann–Whitney U-test (a, b) or two-way ANOVA (c–f). P-values are shown in the graphs. AUC area under the curve, MFI mean fluorescence intensity, OC osteoclast