Research Paper Volume 13, Issue 7 pp 10312—10325

Membrane-cytoplasm translocation of annexin A4 is involved in the metastasis of colorectal carcinoma

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).

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).