Extended Data Fig. 10: Additional analyses of the activity-dependent splicing of Lphn3 Exon31 and Exon32.

a, Neuronal activity does not change the Nrxn1 SS4 splicing level, as reported before³⁶. This serves as a negative control to ensure that the strong activation conditions (KCl and kainate) used did not impair neuronal survival. Two-sided t test was used to calculate statistical significance showing only significant p < 0.05. n = 3 biologically independent cortical cultures; n = 8, 7, 10 biologically independent hippocampal tissues after 0 hr, 2 hr, and 10 hr Kainate treatment, respectively. b, UMAP plot illustrating the clustering of neurons (NeuN⁺) into subtypes of excitatory and inhibitory groups in a single-cell RNAseq dataset obtained from the mPFC³⁷. N, neuron; Ex, excitatory; In, inhibitory. Scl30a3, Rprm, Tshz2 are markers identifying distinct groups of excitatory neurons, while Vip, Sncg, Lhx6 and Reln are markers identifying distinct groups of inhibitory neurons. c, Expression levels of the indicated marker genes in different types of excitatory and inhibitory neurons in the mPFC single-cell RNaseq dataset. d, Mapping of the expression levels of immediate early genes (IEG) as activation markers onto the neuron types identified in the UMAP plot of b. e, Total Lphn3 levels determined in the various RNAseq datasets used for the analysis of activity-dependent alternative splicing of Lphn3. Two-sided Wilcoxon rank-sum test was used to calculate statistical significance showing only significant p < 0.05. n = 3 biologically independent cortical cultures; n = 4 biologically independent hippocampal cultures; n = 8, 7, 10 biologically independent hippocampal tissues after 0 hr, 2 hr, 10 hr Kainate treatment, respectively; the single cell study analysed n = 903, 1526, 232 independent cells for IEG low, medium, and high, respectively.