(A) Bone marrow-derived macrophages (BMDMs) were incubated overnight with conditioned media (CM) from BCL-xL dependent SVEC cells that had been treated with ABT-737 (10 μM) – CM (APO)- or ABT-737 (10 μM) and Q-VD-OPh (30 μM) – CM (CICD)- for 8 h. Expression of indicated M1 and M2 markers was determined by qRT-PCR.
(B) BMDMs were treated as in (A) and stained for either M1 (CD86+) or M2 (CD206+) surface markers. n=3 independent experiments; mean values ± S.E.M.
(C) BMDMs were incubated overnight with conditioned media (CM) from vector or APAF-1 knockdown BCL-xL dependent SVEC cells that had been treated with ABT-737 (10 μM) for 8 h. qRT-PCR was performed on BMDMs to assess M1 marker expression.
(D) BMDMs were incubated overnight with conditioned media (CM) from control or BAX/BAK deleted BCL-xL dependent SVEC cells that had been treated with ABT-737 (10 μM) and Q-VD-OPh (30 μM) for 8 h. qRT-PCR was performed on BMDMs to assess M1 marker expression.
(E) BMDMs were incubated overnight with conditioned media (CM) from vector or IκBSR overexpressing BCL-xL dependent SVEC cells that had been treated with ABT-737 (10 μM) and Q-VD-OPh (30 μM) for 8 h. qRT-PCR was performed on BMDMs to assess M1 marker expression.
(F) BMDMs were incubated overnight with conditioned media (CM) from murine primary pancreatic tumour cells (Myc-PDAC) that had been treated for 24 h with ABT-737 (10 μM) +/- Q-VD-OPh (10 μM). M1/M2 status was assessed with qRT-PCR.
(G) BMDMs were incubated overnight with conditioned media (CM) transferred from murine primary pancreatic tumour cells (Myc-PDAC) that have been treated for 24 h with ABT-737 (10 μM) +/- Q-VD-OPh (10 μM). The activation profile was measured by FACS, staining for either M1 surface marker (CD86+) or M2 surface marker (CD206+). n=3 independent experiments; mean values ± S.E.M. For (A)(C)(D)(E)(F) data represent the mean of triplicate samples and are representative of three independent experiments.
*p<0.05, **p<0.01, ***P<0.001; Tukey-corrected one way ANOVA (B, G). Statistical source data can be found in Supplementary Table 5.