CD40 activation induces transmigration of MM.1S MM cells.
(A) Serum-starved MM.1S cells were plated on a polycarbonate membrane (8-μm pore size) in the transwell cluster plate and activated using 0 to 50 μg/mL of anti-CD40 mAb (G28.5 or 626.1) or sCD40L added to the lower chamber. After 6 hours of incubation, migrating cells in the lower chamber were counted. G28.5 (■); 626.1 (▪); sCD40L (▧). (B) Serum-starved MM.1S cells were plated at the upper chamber in the transwell cluster plate and activated using 0 to 10 μg/mL G28.5 anti-CD40 mAb added in the lower chamber (■) or added both in the upper and lower chambers (▪). Migrating cells into the lower chamber were collected and quantitated. (C) Membranes separating upper from lower chambers in the transwell were coated with (▪) or without (■) fibronectin (40 μg/mL) overnight. Results are mean ±SE of 3 independent experiments. (D) MM.1S were incubated with CD40 stimulants (μg/mL): G28.5 (■); 626.1 (▪); sCD40L (▧). After 36 hours, the cells were pulsed with 3H-thymidine, and DNA synthesis was measured. Results are mean ±SE of 3 independent experiments.
