Figure A2.

(A) EDEM3 constructs are processed by EndoH. EDEM3-KO cells were transfected with plasmids encoding for EDEM3 constructs. Lysates were treated with EndoH and subjected to Western blotting. (B) Same as in (A), except lysates were also incubated with PNG-aseF. (C) Western bloting analysis of NHK processing by EDEM3 constructs in HEK293T cell line. After co-transfection with NHK and EDEM3 constructs, cells were lysed and processed for WB. Rabbit anti-A1AT and rat anti-HA antibodies were used for detection. Actin was used as loading control. (D) NHK intracellular processing in EDEM3-KO and MAN1B1-OE cells. Following co-transfection with the appropriate plasmids, cells were starved, pulse labeled and chased for the indicated time points. Immunoprecipitation was performed using rabbit anti-A1AT antibodies. Resulting immunocomplexes were separated by SDS-PAGE and results were visualized by autoradiography. (E) Same as in (D) with the exception that ST was used.