Extended Data Fig. 1: O-GlcNAcylation regulates angiogenic transdifferentiation by H3.3 deposition.

a. Western blots confirm the overexpression of SNAP-tagging H3.3 in Hela cells. The fusion protein SNAP-H3.3 is detected by the SNAP antibody at 37 kD and SNAP itself at 25 kD. Western blot for H3.3 shows endogenous H3.3 at 15 kD and SNAP-H3.3 at 37 kD. b. Immunofluorescence imaging of positive (P = pulse), negative (QP = quench-pulse), and experimental (QCP = quench-chase-pulse) controls for the quench-chase-pulse experiments (Scale bar, 15 µm). c. Quantification of immunofluorescence from quench-chase-pulse controls. P: n = 126 cells; QP: n = 162 cells; QCP: n = 119 cells. P values are from Kruskal-Wallis test. d. Total protein visualization for indicated groups on gel using Stain-Free Technology. e. Confirmation of HIRA knockdown efficiency in BJ fibroblasts by western blot. f. Putative site-specific O-GlcNAcylation site mutations on HIRA, confirmed by Sanger sequencing. g. Verification of HIRA WT and HIRA mutant overexpression in BJ fibroblasts through Western blot using a Flag-tag antibody. In c, data are mean ± SD. In a, e, g, results are representative of two independent experiments. In d, results are representative of three independent experiments.