Fig. 4.

Insulin promotes Ca²⁺ flux in iPSC-derived BFCNs. Cells cultured in 96-well plates were insulin deprived overnight before insulin or vehicle was added to a final concentration of 1000 ng/ml after 25 s of baseline recording (data from a total of 4 experiments). a Calcium response to insulin normalized to 20 s of baseline recording before insulin addition. b Calcium response to addition of vehicle recorded as arbitrary relative fluorescent units. c Visual corroboration and quantification of Ca²⁺ in individual cells. d Average normalized calcium responses for baseline, (−) Insulin, and response, (+) insulin, in IPSC-derived basal cholinergic neurons. An effect by insulin exposure was detected by 2-way ANOVA (p < .0001). Bonferroni comparisons ***, P < .001; ** P < 0.01