Fig. 7.

C2-ceramide and SH-BC-893 promote actin polymerization. A, FL5.12 cells treated with SH-BC-893 (10 μm) or C2-ceramide (50 μm) for 1 h and then stained for F-actin with Alexa Fluor 488-conjugated phalloidin. B, Same as in A, except cells pre-treated with vehicle or PP2A inhibitors calyculinA (calyA, 5 nm) or LB-100 (100 μm) for 1.5 h before SH-BC-893 addition. C, Same as in A, except cells pre-treated with vehicle or actin polymerization inhibitor latrunculinA (LatA, 1 μm) for 30 min before SH-BC-893 (2.5 μm) or C2-ceramide (30 μm) addition. D, FL5.12 cells pre-treated (30 min) with LatA (1 μm) were treated with SH-BC-893 (2.5 or 5 μm) or C2-ceramide (30 or 50 μm) for 1 h and surface CD98 quantified by flow cytometry. E, FL5.12 cells pre-treated with LatA (30 min, 1 μm) were treated with SH-BC-893 (5 μm) for 2 h and imaged by DIC microscopy to visualize vacuoles. n = 3 in D and n ≥ 30 cells per condition in E. A two-tailed t test was used to compare SH-BC-893 -treated cells to combination of LatA and SH-BC-893, *** = p ≤ 0.001.