Fig. 4: FGF signalling mediates non-cell autonomous upregulation of BCL-2 proteins.

a A growth factor membrane ligand array was probed with supernatant from control or venetoclax treated HeLa tBID2A cells. Spots for FGF2 are indicated. b Levels of FGF2 were determined in supernatant from control or venetoclax treated HeLa tBID2A cells by ELISA. n = 3 independent experiments; mean values ± s.e.m.; unpaired, two-sided t-test. c HeLa tBID2A cells were treated with recombinant FGF2, harvested after 6 h and protein expression analysed by western blot (representative blot of three independent repeats). d Control or FGF2^CRISPR HeLa tBID2A cells were directly treated with venetoclax or the respective supernatant for 48 h as indicated before harvesting and analysis of protein expression by western blot (representative blot of three independent repeats). e Supernatant from untreated or venetoclax treated HeLa tBID2A cells was supplemented with 500 nM trametinib before addition onto recipient cells. After 3 h, recipient cells were harvested and RNA expression of FGF target genes analysed by RT-qPCR. n = 4 independent experiments; mean values ± s.e.m.; *p < 0.0001 compared to SN Ven; Dunnetts corrected one-way ANOVA. f Supernatant from control or venetoclax treated HeLa tBID2A cells was supplemented with decreasing doses of FGFR inhibitors as indicated (AZD: 5 µM, 2.5 µM, 1.25 µM; PRN1371: 10 µM, 5 µM, 2.5 µM) before addition onto recipient cells for 48 h and analysis of protein expression by western blot (representative blot of three independent repeats). g HeLa tBID2A cells were transfected with siRNA targeting FGFR1 and FGFR3 alone or in combination for 24 h before addition of 500 nM venetoclax, harvesting after 48 h and analysis of protein expression by western blot (representative blot of three independent repeats). h HeLa tBID2A cells were transfected with siRNA targeting FGFR1 and FGFR3 alone or in combination for 24 h before addition of control or venetoclax treated supernatant from control cells, harvesting after 48 h and analysis of protein expression by western blot (representative blot of three independent repeats).