could not tell the identity of the subject but could tell which two lines belonged to the same subject, so that the person performing the experiments could use at least one line for each subject without knowing the group category. The collected data were used for statistical analysis without exclusion. All experiments were performed in technical and biological triplicate and were repeated at least three times. The variation within each group of neurons was not pre-estimated and the variation between groups might not be similar. For electrophysiological recording experiments, at least five to six neurons per subject were recorded and statistically analysed without exclusion. For Ca2+ imaging and immunostaining experiments, typically four or five view fields per subject containing several hundred neurons were used without exclusion for analysis. For RNA-seq, qRT–PCR and flow cytometry experiments, all cells in one culture were collected for analysis.
