Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2011 Jan 1.

Published in final edited form as: Exp Cell Res. 2009 Sep 19;316(1):92–102. doi: 10.1016/j.yexcr.2009.09.015

(A) Schematic representation of full length and NLS deleted MORF4 (MORF4Flag ΔNLS). (B) Stability of MORF4Flag ΔNLS mutant in HeLa cells. Full length MORF4Flag and MORF4Flag ΔNLS constructs were transiently transfected into HeLa cells and the cells incubated overnight. Two hours after MG132 treatment, total cell lysates were prepared and MORF4 protein levels were detected by Western blot using mouse anti-Flag (M2) antibody. The location of molecular markers is indicated (kDa). C; cyctoplasmic fraction and N; nuclear fraction. (C) Ubiquitinated MORF4 is predominantly localized in the cytoplasm in HeLa cells. MORF4Flag and MORF4Flag ΔNLS constructs were transiently transfected into HeLa cells with and without HA-Ub and the cells incubated overnight. Twenty-four hours after transfection, cytoplasmic and nuclear lysates were prepared and MORF4Flag and MORF4Flag ΔNLS protein in each fraction was immunoprecipitated with anti-Flag (M2) antibody. Ubiquitinated proteins were detected by Western blot with rabbit anti-HA antibody. Ten percent of each lysate was used as an Input to confirm fractionation. The location of molecular markers is indicated (kDa). C; cyctoplasmic fraction and N; nuclear fraction. (D) Localization of MORF4Flag ΔNLS protein in HeLa cells. Full length MORF4Flag and MORF4Flag ΔNLS constructs were transiently transfected into HeLa cells and the cells were incubated overnight. Two hours after MG132 treatment, MORF4Flag protein was detected by immunocytochemistry using anti-Flag (M2) antibody.

(A) Schematic representation of full length and NLS deleted MORF4 (MORF4Flag ΔNLS). (B) Stability of MORF4Flag ΔNLS mutant in HeLa cells. Full length MORF4Flag and MORF4Flag ΔNLS constructs were transiently transfected into HeLa cells and the cells incubated overnight. Two hours after MG132 treatment, total cell lysates were prepared and MORF4 protein levels were detected by Western blot using mouse anti-Flag (M2) antibody. The location of molecular markers is indicated (kDa). C; cyctoplasmic fraction and N; nuclear fraction. (C) Ubiquitinated MORF4 is predominantly localized in the cytoplasm in HeLa cells. MORF4Flag and MORF4Flag ΔNLS constructs were transiently transfected into HeLa cells with and without HA-Ub and the cells incubated overnight. Twenty-four hours after transfection, cytoplasmic and nuclear lysates were prepared and MORF4Flag and MORF4Flag ΔNLS protein in each fraction was immunoprecipitated with anti-Flag (M2) antibody. Ubiquitinated proteins were detected by Western blot with rabbit anti-HA antibody. Ten percent of each lysate was used as an Input to confirm fractionation. The location of molecular markers is indicated (kDa). C; cyctoplasmic fraction and N; nuclear fraction. (D) Localization of MORF4Flag ΔNLS protein in HeLa cells. Full length MORF4Flag and MORF4Flag ΔNLS constructs were transiently transfected into HeLa cells and the cells were incubated overnight. Two hours after MG132 treatment, MORF4Flag protein was detected by immunocytochemistry using anti-Flag (M2) antibody.