Fig. 3: Projection-TAGs revealed axonal collaterals and complex wiring diagram.

a Bar plot showing the percentage of snRNA-seq nuclei, grouped by the number of unique Projection-TAGs detected in each nucleus. b Heatmap demonstrating the pairwise overlapping of snRNA-seq nuclei positive for any two Projection-TAGs, shown as the percentage of nuclei positive for target 1 TAG (column) that are also positive for target 2 TAG (row). c, d Heatmaps showing the Projection-TAG (row) detected in individual snRNA-seq nuclei (column) expressing only one Projection-TAG (c) and multiple Projection-TAGs (d). Nuclei are ordered based on the projection patterns, and only groups that passed the FDR cutoff were shown on the heatmaps. e Venn diagram showing the overlap between VP-TAG-expressing and other ET-TAG-expressing snRNA-seq nuclei. Other ET-TAGs include Projection-TAGs injected into PAG, MY, SC_L, and SC_S. f Representative FISH image showing MOp cells labeled with VP- and other ET-TAGs. Scale bar = 100 µm. Experiments were repeated in three mice. g Distribution of transcriptional subtypes in each projection group. Transcriptional subtypes with <60 nuclei from each projection feature group were excluded, and cell types that make up at least 10% nuclei in the projection group are annotated on the plot. h Heatmap showing the z-score of average expression of 118 genes (row) differentially expressed in transcriptional L5 PT nuclei in each projection group (column). Data shown on the heatmap can be found in Supplementary Data 5.