Fig. 4: Pyruvate supplementation ameliorates neuronal phenotypes caused by SETD1A haploinsufficiency.

A Representative IF images of neurite outgrowth from cortical spheroids after 48 h of static culture in media supplemented with either 1 mM pyruvate, 1 mM lactate, or 0.5 mM glucose. B Quantification of neurite outgrowth in response to metabolite supplementation (n = 6 spheroids per line per treatment, from 3 independent differentiations. C Representative calcium imaging traces of ΔF/F₀ over time in active neurons after 2 weeks of metabolite supplementation. D Mean spike rate of active neurons within a neuronal network after 2 weeks of metabolite supplementation (n = 6 wells per line per treatment, from 3 independent differentiations). E Cumulative fraction of neurons by spike frequency (n = 213 (WT/no treatment), 202 (WT/pyruvate), 208 (WT/lactate); 217 (Clone 1/no treatment), 162 (Clone 1/pyruvate), 155 (Clone 1/lactate); 153 (Clone 2/no treatment), 183 (Clone 2/pyruvate), 100 (Clone 2/lactate) neurons recorded, from 3 independent differentiations). Data represented are mean ± sd. For all comparisons, two-way ANOVA with Sidak’s multiple comparisons test was performed. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.