Fig. (1) Ubiquitylation of Sec23a. Huh7 cells were co-transfected with the pHA-Ubiquitin expression vector and the 3xFLAG-Sec23a vector for 48 h and then subjected to cycloheximide treatment for the indicated time. Cell lysates were immunoprecipitated by a mouse monoclonal anti-HA antibody to selectively pull down ubiquitinated proteins and then the 3xFLAG-Sec23a was revealed by a mouse monoclonal anti-FLAG antibody. Total lysates were checked by Western Blot with the indicated antibodies as control of the input samples.