Fig. 5

Claustrum cells in neonatal mice display the same Nurr1/Tle4 expression pattern as adult mice. A Injection of retrograde AAV-CAG-TdT into the anterior cingulate cortex (ACC) at different postnatal days, followed by tissue collection 7 days post injection. B1–B4 Examples of AAV injection sites in the ACC at P7 (B1), P14 (B2), P21 (B3) and P56 (B4) following injection at P1, P7, P14 and P49, respectively. C–F Representative images C, E showing the colocalization of TdTomato (TdT) labeling with Nurr1 (C) and Tle4 (E) in the anterior claustrum (CLA) at P7 (left, merged imaging channels; right, single channel images). Venn diagrams D, F representing the mean number of cells at P7 expressing Nurr1 and TdT (D), or Tle4 and TdT (F) (n = 5 mice). Values shown are the average cell counts from the anterior, middle and posterior coronal planes of the CLA. G–J Same as C–F, but for mice injected at P7 and perfused at P14 (n = 6 mice). K–N Same as C–F but for mice injected at P14 and perfused at P21 (n = 7 mice). O–R Same as C–F, but for mice injected at P49 and perfused at P56 (n = 5 mice). S Quantification of fluorescence intensity in the CLA region (cyan) in the medial–lateral axis (dashed line). T Measurement of fluorescence intensity (z-score) for TdT (cyan), Nurr1 (magenta) and Tle4 (black) labeling across the mediolateral axis of the CLA region at P7, P14, P21 and P56 (from left to right). Solid lines for each color represent the mean and light shaded areas of the same color represent standard deviation. U, V: Same as S–T, but for the dorsoventral axis. Insets in left panels of C, E, G, I, K, M, O, Q (bottom-left) are 2.5 × fold magnifications of areas in white boxes. Orange arrowheads indicate examples of cell colocalization. Values in D, F, H, J, L, N, P, R represent mean ± standard deviation (see Additional file 2: Table S3 for details on mouse sex)