Supplementary Figure 3: Axon paths and marker labelling of Opn5 lineage cells.

a-c, tdTomato signal (red) in Opn5^cre; Ai14 P28 mouse brain cryosections in the optic tracts (a), superior colliculus (b) and lateral geniculate nucleus (c). These projections are characteristic of retinal ganglion cells. d-g, Opn5^cre; Ai6 P8 retinal cryosections imaged for nuclei with DAPI (d, grayscale), for ZsGreen1 from Ai6 (d, e, green), for RBPMS (d, f, red) and for Calretinin (d, g, blue). White outlines of green cells are displayed on the red RBPMS (f) and blue Calretinin (g) channels. in (d) a displaced RGC and some of the amacrine cells are indicated. RBPMS is exclusive to RGCs and Calretinin identifies both amacrine cells and a subset of RGCs. We determined the RBPMS and calretinin status of Opn5^cre; Ai6 positive cells from n-3 mice at P8. White outlines of Opn5^cre; Ai14 positive cell bodies were moved to images of the RBMPS and Calretinin labelling channels and counts performed. In these sections there were 88 Opn5^cre; Ai6 positive cells. In these same sections, there were 1446 RBPMS positive cells and 910 Calretinin positive cells. All 88 Opn5^cre; Ai6 positive cells were positive for RBPMS and 50/88 were also positive for Calretinin. Importantly, there were zero Opn5^cre; Ai14 positive cells that expressed only calretinin; they invariably also expressed RBPMS. This analysis provides evidence that Opn5^cre is expressed is RBPMS positive RGCs, but not in amacrine cells. Scale bars 500 μm (a) 200 μm (b, c) 50 μm (d). Image panels are representative of at least three separate experiments.