Figure 4.Figure 4
Direct effects of acetylcholine on DMG (A) Timeline for the generation of iPSC-derived cholinergic with viral transduction (AAV-VAChTe1-ChRmine::eYFP or AAV-VAChTe1::eYFP) for co-culturing with glioma cells. (B) Confocal micrographs showing cholinergic motor neurons generated hiPSCs of a healthy 12-year-old male and co-cultured with DMG cells of an 8-year old male. NF-H/M, turquoise; H3K27M, white; scale bars, 50 μm. (C) Quantification of glioma cell proliferation (EdU + /H3K27M + ) when co-cultured with hiPSC-derived cholinergic motor neurons paired with in vitro optogenetic stimulation. One-way analysis of variance (ANOVA) with Tukey’s post hoc analysis; **** p < 0.0001, ** p < 0.01, ns: non-significant. Data = mean ± SEM; n = four independent experiments, each with three wells per condition; each data point represents the mean of three wells per condition for a given experiment. (D) Three-dimensional rendering illustrating presynaptic cholinergic neuron (NF-H/M: white) with pr