Figure 7.

Streptolysin-O permeabilizes Lec35 cells and preserves the mannosylation defect. CHO-K1 (A and B), Lec35.1 (C and D), and Lec15.2 (E and F) cells were treated with SLO either when attached to dishes (Monolayer, A, C, and E) or after suspension (Suspended Cells, B, D, and F). The permeabilized cells were incubated with either 2.0 ml (A, C, and E) or 0.2 ml (B, D, and F) of transport buffer (Martys et al., 1995) at 37°C for 10 min containing 1 μM unlabeled UDP-GlcNAc to initiate LLO synthesis, 0.1 μCi GDP-[³H]mannose to label and extend LLOs, and 0.2 mM 5′ AMP to inhibit breakdown of nucleotide sugars. LLO were recovered and characterized by HPLC.