Fig. 6

Effects of avapritinib, alisertib and bortezomib in SM cell lines. A Induction of apoptosis by alisertib 250 (blue) and 500 (orange) nM used as single agent after 24 and 48 h of treatment in HMC-1.1, -1.2 and in ROSA^{KIT D816V} cells. Columns represent the mean of three independent cytofluorimetric assays performed by loading cells with Annexin V (FITC) and PI and bars represent the standard error. B Induction of apoptosis by alisertib (500 nM) or bortezomib (10 nM) alone and in combination with avapritinib (250 nM) after 24 (lblue) and 48 (orange) hours of treatment in HMC-1.2 and ROSA^{KIT D816V} cells. Columns represent the mean of three independent cytofluorimetric assays performed by loading cells with Annexin V (FITC) and PI and bars represent the standard error. C Avapritinib (250 nM, orange), alisertib (500 nM, grey) and bortezomib (10 nM, yellow) administration, alone and in combination (AVA + ALIS, light blue; AVA + BORT, green) in HMC-1.2 and ROSA^{KIT D816V} cells maintained in liquid medium and counted every 24 h up to 72 h of treatment showed that the combinations did not significantly potentiate the cytotoxic effects of either drug alone. Each point represents the mean of three independent counts performed after trypan blue staining by using the burker chamber. D Induction of cell death by avapritinib (250 nM) and alisertib (500 nM) or bortezomib (10 nM) treatment in HMC-1.2 cells assessed by using an EVOS XL Core optical microscope equipped with a 20 × objective. E Avapritinib (100 nM, red), alisertib (200 nM, green) and bortezomib (2 nM, yellow) administration, alone and in combination (AVA + ALIS, light blue; AVA + BORT, orange) in HMC-1.2 cells maintained in liquid medium and counted every 24 h up to 72 h of treatment showed that the combinations, at low doses of each drug, significantly potentiate the cytotoxic effects of either drug alone. Each point represents the mean of three independent counts performed after trypan blue staining by using the burker chamber