Fig. 6: In vitro validation identifies opposing effects of in silico drug predictions and top schizophrenia eGenes (related to Box 2).

In vitro validation of drug-eGene phenotypic interactions. A Effects of 48-hour treatment with 10 µM simvastatin on synaptic puncta density in TMEM219 CRISPRa perturbed (teal) or non-perturbed (purple) iGLUT neurons. Syn1-positive puncta values are expressed relative to MAP2-positive neurite length in each well. Perturbation of TMEM219 expression with CRISPRa significantly increased synaptic puncta density; this increase was partially ameliorated by 48 hr treatment with 10 µM simvastatin (two-way ANOVA; CRISPRa variation p < 0.0001; CRISPRa x Drug treatment variation p < 0.05). N minimum of two independent experiments across 2 donor lines with 12 technical replicates per condition. Values for each technical replicate in imaging experiments were averaged from nine separate images per single well. B Treatment of cells perturbed with either TMEM219 CRISPRa with 10 µM Simvastatin reverses or suppresses the transcriptomic impacts of the schizophrenia eGene perturbations alone (SI Figs. 28–30). Treatment of cells with CRISPRa TMEM219-gRNA and 10 µM Simvastatin over 48 hours opposes the transcriptomic impact observed in CRISPRa TMEM219-gRNA + Vehicle-treated cells. Venn diagram of significant DEGs at an (top left) adjusted p val ≤ 0.05 and at an (top right) unadjusted p value of ≤0.05. (bottom) Dot plot demonstrating the logFC of each gene in either the TMEM219 + Vehicle (green) or TMEM219 + 10uM Simvastatin (yellow) condition, ordered by degree of logFC in the TMEM219 + Vehicle-treated cells. Size of the points corresponds to the −log10 (adjusted p value).