Figure 2.

The human UCE expressed by stably transfected L-cells is sialylated. Membrane extracts of cells expressing human UCE wild type (lanes 1 and 2), Y488A mutant (lanes 3 and 4), or H510 stop mutant (lanes 5 and 6) were treated (+; lanes 2, 4, and 6) or not treated (−; lanes 1, 3, and 5) with Vibrio cholera neuraminidase (Neur.) for 80 min at 37°C. The reaction mixtures were boiled in SDS-PAGE sample buffer and subjected to reducing SDS-PAGE on a 7.5% gel. The gel was blotted to nitrocellulose, which was probed with affinity-purified rabbit anti-human UCE antibody (1:1000) and detected by enhanced chemiluminescence.