FIGURE 3.

A, Western blot analysis of C-HA-tagged WT-PCFT, H247A, H281A, and H312A mutants. B, Western blot analysis of WT- and the N58Q/N68Q- PCFT [deglycosylated], PCFT along with WT- and H247A-PCFT proteins after tunicamycin treatment (1 μg/ml) of HeLa cells for 48 h. In both panels, equal amounts of crude cell plasma membrane protein fractions (30 μg) were loaded, and the blots were probed with an anti-HA antibody. The numbers on the left indicate the molecular sizes in the protein ladders. The lower bands in each panel indicate the β-actin loading control. The blot is representative of three (A) and two (B) separate experiments.