Fig. 4

DLEU1 inhibits RNF114-mediated ubiquitination of DYNLL1. A, B KYSE-150-DLEU1 cells transfected with Flag-tagged DYNLL1 and Myc-tagged DYNLL1 and treated with MG132 were immunoprecipitated with anti-Flag (A) or anti-Myc (B) antibody followed by immunoblotting with anti-Myc or anti-Flag antibody. C Lysates from KYSE-150-DLEU1 cells transfected with the indicated expression constructs and treated with MG132 were subjected to immunoprecipitation with anti-Flag antibody followed by immunoblotting with ubiquitin antibody. D Lysates from KYSE-150-DLEU1 cells transfected with the indicated expression constructs and treated with MG132 were subjected to immunoprecipitation with anti-DYNLL1 antibody followed by immunoblotting with ubiquitin antibody. E Lysates from KYSE-410 cells transfected with siRNF114 and/or shDLEU1 were subjected to immunoblotting with a DYNLL1 antibody. β-actin was used as a loading control. F Lysates from KYSE-150 cells transfected with Myc-RNF114 and/or DLEU1 were subjected to immunoblotting with anti-DYNLL1. G Sequences of wild-type and mutated DYNLL1. H Lysates from KYSE-150 cells transfected with the indicated expression constructs and treated with MG132 were immunoprecipitated with anti-Flag antibody followed by immunoblotting with ubiquitin antibody. I Lysates from KYSE-150 cells transfected with the indicated expression constructs were subjected to immunoblotting analysis. β-actin served as a loading control