The stability of WBSCR22 protein is regulated by TRMT112.
(A) The expression level of WBSCR22 and its interaction partners in response to siRNA treatment was analyzed by western blot in HeLa cells. The cells electroporated with siRNAs were collected 48 h after transfection, lysed and analyzed with antibodies against WBSCR22, TRMT112, C1QBP and α-tubulin. (B-D) TRMT112 is required for WBSCR22 stability in HeLa (B), U2OS (C) and COS-7 (D) cells. Cells were treated with TRMT112 siRNAs and analyzed by antibodies against WBSCR22, TRMT112 and α-tubulin. (E) Expression of TRMT112 restores the endogenous WBSCR22 protein level in TRMT112-depleted U2OS cells. First, the U2OS cells were electroporated with siRNAs and 48 hours later with the plasmid encoding for E2Tag-TRMT112. Cells were collected 24 hours after second transfection and analyzed by western blotting with antibodies against WBSCR22, TRMT112 and α-tubulin. Proteins from 105 cells were loaded on each lane.
