Fig. 3
A starvation microenvironment promotes proliferation, invasion, and migration by facilitating STAM2 O-GlcNAcylation. (A) STAM2 IP assay was conducted in UM-UC-3 cells transfected with either STAM2-HA or vector. STAM2-HA and O-GlcNAc were detected using the indicated antibodies. (B-C) Co-IP assays validated the interaction between STAM2-HA and OGT-Flag using anti-HA (B) or anti-Flag (C) antibodies. (D) STAM2 IP assay in UM-UC-3 cells transfected with STAM2-HA or vector (HBSS incubation for 12 h). (E) Quantitative western blot analysis of STAM2 O-GlcNAcylation. (F-G) Proliferation, migration, and invasion were measured using CCK-8 and Transwell assays. (H) EMT marker protein levels were measured using western blot. (I) Quantitative western blot analysis of H. (J) STAM2 O-GlcNAcylation was detected via an IP assay in UM-UC-3 cells transfected with STAM2-HA, STAM2 S372-HA, or STAM2 S375-HA. (K) Quantitative western blot analysis of STAM2 O-GlcNAcylation. (L-M) Proliferation, migration, and invasion were measured using CCK-8 and Transwell assays. (N) EMT marker protein levels were measured using western blotting. (O) Quantitative western blot analysis of N. All PVDF bands were transferred and tested in the same western blot experiments. All the results are based on three distinct repetitions. Statistical significance is denoted as ***p < 0.001, **p < 0.01, and *p < 0.05, indicating significant differences between two groups.
