Extended Data Fig. 3: Activity-dependent gene expression patterns detected by total RNAseq.

a. mRNA expression level changes measured by total RNAseq of 4 G hGNs at 15 minutes (n = 4), 1 hour (n = 3), 2 hours (n = 4), and 4 hours (n = 4) after membrane depolarization compared to expression in unstimulated cultures (n = 4) represented by MA-plot. Genes with a significantly different gene expression level and a minimum fold-change magnitude of 1.5 after depolarization are marked in red. Genes having inducible expression fall above y=0 and example gene names are labeled. b. Time courses of previously known activity-inducible gene transcript levels in unstimulated and stimulated 1 G hGNs. Means±s.e.m. Timepoints at which the transcript was significantly induced compared to unstimulated cultures are marked with an asterisk. c. qPCR Fold-induction of mRNA transcripts in 1 G hGNs cultures of genes detected to be inducible by total and single cell RNAseq. Sensitivity limitations of qPCR prevent detection of significant differences when a gene’s expression is modestly induced in a minority of the cells (for example MMP1 and SHANK3). d. The percentage of 1 G and 4 G hGN inducible genes whose gene products have known subcellular localizations. Early inducible genes (15 min. - 1 hr.) have a greater percentage of nuclear localized products than gene transcripts induced at later stimulation timepoints, when the percentage of induced genes in the membrane-bound or secreted category increases.