![Fig. 5. CD40 induces migration in human MM cells expressing mutant p53. / MM cells were plated on a polycarbonate membrane (8 μm pore size) precoated with fibronectin (5 μg/mL [μM]) in the transwell cluster plate and were activated using 0 to 10 μg/mL (μM) sCD40L (A) or anti-CD40 mAb (G28.5) (B) added to the lower chamber. Cells were cultured at restrictive temperature (37°C) with mt p53 or at permissive temperature (28°C) with wt p53. Migrated cells in the lower chamber were counted using a Coulter counter ZBII. (C) Isotype control antibody MOPC 21 (10 μg/mL [μM]) did not induce transmigration in MM cells at restrictive temperature. Results are representative of 2 independent experiments. (■) RPMI 8226 at 37°C; (▤) RPMI 8226 at 30°C; (▪) SV at 37°C; () SV at 30°C. (D) Patient MM1-5 cells were subjected to transmigration assay, using sCD40L (10 μg/mL [μM]) in the lower chamber at 37°C. (▪) MM1; () MM2; (■) MM3; (░) MM4; () MM5. Increased transmigration is observed in MM cells expressing mutated p53 (MM2, MM4, MM5), but not in those with wt p53 (MM1, MM3), after CD40 activation.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/99/4/10.1182_blood.v99.4.1419/6/h80422116005.jpeg?Expires=1723753259&Signature=4VwUWs3NBWjNqqkfMD8wGFBElhLfYtcQZ-HKRJc3dwyVNnuaui8ZUtDdbavKK7LIAoDsx569Pme-q-0Le4CRUsdxzUtwcSTZ04HT~JgxHnxfDKsWjWNDzWhXcW-V~ynYkK67jdJcCtX8BeOkM46oZYVxWhZ9ieeOTcNu~eT-bQWLjIzylI6ic0aj2N2MJpG2W~DCWTMwBO1xXvUG34rvk~3qN3aLu9DXHhNNaWabbkVsx5Bf0Z8uQoAOU2ICQK9gexlDP6LEk60rlzIFaek4GyDECAc2d9xwZoV8dVweqtBgJlbXpNxh5uahe4zdwFaxOEGGe9CnO-mY9Di7UMMfNA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
CD40 induces migration in human MM cells expressing mutant p53.
MM cells were plated on a polycarbonate membrane (8 μm pore size) precoated with fibronectin (5 μg/mL [μM]) in the transwell cluster plate and were activated using 0 to 10 μg/mL (μM) sCD40L (A) or anti-CD40 mAb (G28.5) (B) added to the lower chamber. Cells were cultured at restrictive temperature (37°C) with mt p53 or at permissive temperature (28°C) with wt p53. Migrated cells in the lower chamber were counted using a Coulter counter ZBII. (C) Isotype control antibody MOPC 21 (10 μg/mL [μM]) did not induce transmigration in MM cells at restrictive temperature. Results are representative of 2 independent experiments. (■) RPMI 8226 at 37°C; (▤) RPMI 8226 at 30°C; (▪) SV at 37°C; (
) SV at 30°C. (D) Patient MM1-5 cells were subjected to transmigration assay, using sCD40L (10 μg/mL [μM]) in the lower chamber at 37°C. (▪) MM1; (
) MM2; (■) MM3; (░) MM4; (
) MM5. Increased transmigration is observed in MM cells expressing mutated p53 (MM2, MM4, MM5), but not in those with wt p53 (MM1, MM3), after CD40 activation.
