Figure 6.
Wild-type and truncated TPMV H protein transport kinetics. CHO-K1 cells were transfected with wild-type F and (A) HHis-tag or (B) H 90 or an empty vector (mock) for 24 hours before being starved for 30 minutes and pulsed with 100μCi/ml 35S labeled methionine and cysteine for 1 hour and then chased for up to 4 hours. After the indicated chase period, cell lysates were collected and H was immunoprecipitated and digested without or with EndoH (−/+ lanes, respectively) followed by fractioning on 7.5% SDS-PAGE. (C) Kinetics of complex oligosaccharide acquisition. Vertical axis: percent EndoH resistance of three experiments for wild type H (diamonds) and HΔ90 (squares). Horizontal axis: time of chase (hours).