![Fig. 4. CD40 induces VEGF secretion in a dose- and time-dependent fashion. / (A) RPMI 8226 and SV MM cell lysates (15 μg) at intervals (0-48 hours) after CD40 activation were subjected to 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and were transferred to a polyvinylidene difluoride membrane. VEGF protein was detected with anti-VEGF mAb and was visualized by enhanced chemiluminescence. Anti-α–tubulin mAb was used as a loading control. Upper panel, RPMI 8226; lower panel, SV. Similar results were obtained in 2 independent experiments. (B) Cells were cultured at 37°C in triplicate with 0 to 10 μg/mL (μM) sCD40L (circle) or anti-CD40 mAb G28.5 (triangle). (C) Cells were cultured with sCD40L (10 μg/mL [μM]) (circle), anti-CD40 mAb G28.5 (10 μg/mL [μM]) (triangle), medium alone (diamond), or MOPC-21 isotype control IgG (square) at 37°C. VEGF levels in the supernatants at intervals were determined by ELISA. Data are expressed a means ± SE of culture triplicates. Open symbol, RPMI 8226; solid symbol, SV.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/99/4/10.1182_blood.v99.4.1419/6/h80422116004.jpeg?Expires=1723753595&Signature=lBG3Bq3vGza1qUYXdvaIueeVvJ4-ZeydobARGwHzHbU6BNUodm~TGPLMNKYvriV6Fk3LCYaUMJMRaPGFhiuOk2b9gUTPF6aUL9RNhNUNgJNU9-FaR1vL4iw~rOyxsPYGPRElf57PyZCJRt4RnWRjeHqcCaEYioz2tsl~HvKVfCdn9LWQ5bzL5f8HPr8FjJyKtFF9y0yrY~EsXFlWfDhVs6gavIztsBRxl655EwCP6eFVV~R6xTn3yRXQ4GqL~sJ0AwGNAut-ZhsttmT8~THKqdn2VrKHhXFdrxjZ~bQgmTSJGil4e7kLQ7rJwUjupLm22NQCejisHnyP0Fu9i9mfXg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
CD40 induces VEGF secretion in a dose- and time-dependent fashion.
(A) RPMI 8226 and SV MM cell lysates (15 μg) at intervals (0-48 hours) after CD40 activation were subjected to 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and were transferred to a polyvinylidene difluoride membrane. VEGF protein was detected with anti-VEGF mAb and was visualized by enhanced chemiluminescence. Anti-α–tubulin mAb was used as a loading control. Upper panel, RPMI 8226; lower panel, SV. Similar results were obtained in 2 independent experiments. (B) Cells were cultured at 37°C in triplicate with 0 to 10 μg/mL (μM) sCD40L (circle) or anti-CD40 mAb G28.5 (triangle). (C) Cells were cultured with sCD40L (10 μg/mL [μM]) (circle), anti-CD40 mAb G28.5 (10 μg/mL [μM]) (triangle), medium alone (diamond), or MOPC-21 isotype control IgG (square) at 37°C. VEGF levels in the supernatants at intervals were determined by ELISA. Data are expressed a means ± SE of culture triplicates. Open symbol, RPMI 8226; solid symbol, SV.
