Research Paper Volume 13, Issue 7 pp 10312—10325

Figure 4. SUMOylation of ANXA4 stabilized ANXA4 expression. (A) Immunoprecipitation analysis of ANXA4 SUMOylation after cells were cotransfected with flag-ANXA4 and HA-SUMO1, HA-SUMO2, or HA-SUMO3 and then subjected to western blotting with an anti-HA antibody. (B) Western blot analysis of ANXA4 proteins in HT-29 cells after transfection with HA-sumo1, HA-sumo2 or HA-sumo3. Tubulin was used as the loading control. (C) Immunoprecipitation analysis of ANXA4 SUMOylation after cells were cotransfected with flag-ANXA4 and HA-SUMO1, treated with the Ca²⁺ agonist ionomycin (1 μM or 2 μM) at 14 h and 24 h and then subjected to western blotting with an anti-HA antibody. (D) Western blot analysis of the expression of ANXA4 in HT-29 cells after treatment with cycloheximide with the addition of the Ca²⁺ agonist ionomycin (1 μM).