Fig. 1: Effects of SYK inhibition on GI-NETs cells 2D and 3D culture.

SYK expression was detected in both COLO320DM (A) and in GOT1 (B) cells by immunofluorescence staining (SYK is stained magenta, while nuclei are shown in blue). Effects of SYK inhibition on 2D and 3D NETs cell culture. C, E COLO320DM and GOT1 cells were seeded in 96 well plates and incubated for 72 h with increasing concentration of SYK inhibitor, control cells were treated with a vehicle (DMSO). Cell viability in 2D assays (C) was assessed by crystal violet in three independent experiments with at least six replicates each, and it is expressed as the mean ± SEM. *P < 0.05 vs vehicle cells; ***P < 0.0001 vs vs vehicle cells. ****P < 0.00001 vs vs vehicle cells. Cell viability of 3D cells (E) was measured as luminescent output in three independent experiments with at least two replicates each, and it is expressed as the mean ± SEM **P < 0.01 vs vs vehicle cells, ****P < 0.00001 vs vs vehicle cells, significance was calculated by two-way ANOVA using Dunnett’s multiple comparison test. F Representative images of 3D spheroids at baseline, Day 1, and Day 3 of treatment. D Caspase activity was measured as luminescent output in two independent experiments with two replicates each, and it is expressed as the mean ± SEM. ***P < 0.01 vs vs vehicle cells, ****P < 0.00001 vs vehicle cells, significance was calculated by two-way ANOVA using Dunnett’s multiple comparison test.