Figure 5

PI3Kδ/γ regulate IL-6-induced AKT activation in MM. (a) MM1s and primary MM were incubated with idelalisib (1 μm), CZC24832 (1 μm) and duvelisib (1 μm) for 4 h and then stimulated with IL-6 for 15 min before protein extraction. Samples were then analysed for phospho-AKT and phospho-MAPK response using Western blotting. Blots were re-probed to confirm sample loading. (b) MM1s and RPMI8226 cells were incubated with increasing doses of duvelisib for 6 h after which they were stimulated with and without 100 ng/ml IL-6 for 15 min. Protein was extracted and analysed for phospho-AKT and phospho-MAPK via Western blotting. Total AKT and MAPK were used as loading controls. (c) MM1s cells were transduced with lentivirus targeted to PI3Kδ and PI3Kγ or control shRNA for 72 h and then treated with IL-6 for 15 min. Protein was extracted and analysed for phospho-AKT and phospho-MAPK via Western blotting. Total AKT and MAPK were used as loading controls. (d, e) MM primary cells were incubated with BMSC for 24 h and then treated with idelalisib (1 μm), CZC24832 (1 μm) and duvelisib (1 μm) for 24 h, then assessed for apoptosis by PI/Annexin V staining.