Effects of pharmacological inhibition of signalling pathways in macrophages upon CD44-augmentation of phagocytosis.

Abstract

<p>A) Monocyte-derived macrophages were incubated with CD44 mAb for 30 min prior to addition of dibutyryl cAMP at concentrations shown. Macrophages were then co-incubated with apoptotic targets at 37°C for 30 min prior to assessment of phagocytosis by microscopy. Results shown are mean percentage phagocytosis for 3 independent experiments. B) Monocyte-derived macrophages were pre-incubated with 10 µM LY294002 at 37°C and then co-incubated with CD44 mAb for 30 min prior to addition of CMFDA-labelled apoptotic targets. After 30 min, assessment of phagocytosis was made by flow cytometry. Results shown are mean percentage phagocytosis ± SEM for 5 independent experiments. C)Monocyte-derived macrophages were pre-incubated with 50 nM PD98059 at 37°C and then co-incubated with CD44 mAb for 30 min prior to addition of CMFDA-labelled apoptotic targets. After 30 min, assessment of phagocytosis was made by flow cytometry. Results shown are mean percentage phagocytosis ± SEM for 3 independent experiments. D) Monocyte-derived macrophages were pre-incubated with 25 µM PP2 at 37°C and then co-incubated with CD44 mAb for 30 min prior to addition of CMFDA-labelled apoptotic targets. After 30 min, assessment of phagocytosis was made by flow cytometry. Results shown are mean percentage phagocytosis ± SEM for 4 independent experiments. E) Monocyte-derived macrophages were pre-incubated with 25 µM Genistein at 37°C and then co-incubated with CD44 mAb for 30 min prior to addition of CMFDA-labelled apoptotic targets. After 30 min, assessment of phagocytosis was made by flow cytometry. Results shown are mean percentage phagocytosis ± SEM for 3 independent experiments. In panels B-E there was no statistical difference between the percentage phagocytosis recorded in the presence or absence of pharmacological inhibitor following CD44 mAb treatment.</p