Figure 3. Adult co-expression of Brn3 transcription factors and RBPMS in Brn3c^Cre/WT; Rosa26^tdTomato mice.

Whole-mount immunofluorescence staining with anti-Brn3c and RBPMS (a, b) and Brn3b and Brn3a antibodies (c, d) was performed on adult Brn3c^Cre/WT; Rosa26^tdTomato retinas. For each staining, a larger field is shown (a,c), followed by higher magnification of the insets indicated by stippled lines (b, d). For b, d merged channels are followed by tdTomato, Brn3c and RBPMS (b) or tdTomato, Brn3b and Brn3a (d) single channels. Total number of cells in the GCL is revealed by DAPI, shown in the merged channels. Arrows point to triple positive cells. Retina orientation is indicated. (e, f) Pie charts of marker expressing cells for (a) and (c). Numbers represent total number of quantified cells for each marker. For each staining, four 40x fields were collected from the dorsal, ventral temporal and nasal quadrants of left (e) or right (f) retinas from three Brn3c^Cre/WT; Rosa26^tdTomato adult mice. (g, h) Box-whiskers plot distributions corresponding to e,f. For both pie charts and box plots the “DAPI” numbers represent GCL cells that were only positive for DAPI (so negative for all markers). Box plot conventions are explained in Figure 2. Counted cells, sections and mice for each genotype and antibody combination are provided in Supplementary Table 2. Scale bars in c and d are 50 and 25 μm respectively.