Fig. 7
SETBP1 and FoxP2 co-expressing neurons´ intensities correlate with undirected singing. (a) DAPI (color coded in blue), SETBP1 (color coded in red) and FoxP2 (pseudo color coded in yellow) immunostaining in Area X and merge of all channels, arrow pointing to the left highlights to a SETBP1+/FoxP2 + example, arrow pointing down a SETBP1+/FoxP2- neuron. Orthogonal views of the co-localizing cell are shown for each channel. (b) Box plots showing the mean percentage of zebra finch SETBP1 and FoxP2 co-localizing neurons in Area X in different conditions (US, DS, NS and juvenile). Sample size (number of birds): mean of US = 7, DS = 6, NS = 5 and Juveniles = 5. No statistically significant differences between the experimental conditions were detected, one-way ANOVA, Tukey’s Multiple Comparison Test (supplementary Table 2). (c) Density plots of pixel intensities of individual SETBP1 (orange) and FoxP2 (blue) co-localizing neurons in Area X of undirected singers (US), direct singers (DS), non- singers (NS) and 50 PHDs juvenile non-singers (Juvenile). On the right of the density plots we show correlation plots of intensities of SETBP1 and FoxP2 co-localizing neurons in Area X, undirected singers have a significant correlation (Spearman correlation, spearman r = 0.3149, p = 0.0044), all others did not (DS, spearman r = 0.1833, p = 0.1035; NS, spearman r = 0.2264, p = 0.0820; Juveniles, spearman r=-0.09064, p = 0.4910). We analyzed 60 neurons for NS (n = 3 birds), 80 neurons for US (n = 4 birds), 80 neurons for DS (n = 4 birds), and 60 neurons for juveniles (n = 3 birds). Scale bar = 20 μm.
