Fig. 3

Claustrum neurons projecting to different cortical regions exhibit colocalization with Nurr1 and Nr2f2, but not with Tle4. A Injection of retrograde AAV-CAG-GFP into the anterior cingulate cortex (ACC). Claustrum tissue was processed 14 days post injection. B Example of an AAV injection site in the ACC. C1–C4 Representative images from coronal sections of the anterior claustrum (CLA) relative to the insula (Ins) and the striatum (Str) showing retrograde GFP labeling following AAV injection the ACC (C1), along with the expression of Nurr1 (C2), Nr2f2 (C3), Tle4 (C4), and merged Tle4/GFP (C5). All panels are from the same slice, except Nurr1 which is from an adjacent slice. Dashed ellipses highlight the region of low Tle4 expression. D–F: Representative images (left) and Venn diagrams (right) showing colocalization of GFP labeling with Nurr1 (D), Nr2f2 (E) and Tle4 (F) in the anterior CLA. Panels E and F are the same slice. Venn diagrams in each panel show the mean number of cells expressing GFP with Nurr1 (D), Nr2f2 (E), and Tle4 (F) (n = 5 mice, all male). Values shown are the average cell counts across anterior, middle, and posterior planes of the CLA. Insets in left panels of D, E, F are 2.5 × fold magnifications of areas in white boxes. Orange arrowheads indicate examples of cell colocalization. G–L Same as A–F but for experiments with retrograde AAV-CAG-GFP injected into the primary motor cortex (MOp) (n = 4 mice, all male). M–R Same as A–F, but for retrograde AAV-CAG-GFP injected into the lateral entorhinal cortex (LEC) (n = 4 mice, all male). Values in right panels of D, E, F, J, K, L, P, Q, R represent mean ± standard deviation