Fig. S8.

Melanopsin stimulates mTOR in a light-dependent manner. (A) Whole-mount retinas from intact Opn4-GFP mice housed under a normal 12:12 light–dark cycle (NC) or a 12:12 dark–dark cycle (DD). Retinas were stained with Tuj1 (red) and pS6 (blue) antibodies. GFP (green) labels M1–M3 ipRGCs. (B) Quantification of the percentages of GFP⁺ ipRGCs that express pS6 for the two light conditions. *P < 0.05, Student’s t test, three mice in each group. (C) Whole-mount retinas from intact WT mice housed under a normal 12:12 light–dark cycle (NC) or a 12:12 dark–dark cycle (DD). Retinas were labeled with SMI32 (red) and pS6 (blue) antibodies. (D) Quantification of the percentage of SMI32⁺ αRGCs expressing pS6 for the two light conditions. (E) Quantification of RGC survival for WT mice housed under NC or DD at 2 wk after crush. The mice were injected with AAV-Mela 4 wk before injury. (F) Quantification of the percentage of SMI32⁺ αRGCs expressing pS6 at a different time after continuous light stimulation in injured mice with AAV-Mela or AAV-GFP injection. Mice were dark-adapted for 24 h before stimulation. (G) Sections of optic nerves with CTB-labeled axons from WT mice injected with AAV-shPten and crush. Mice were housed under NC or DD for 2 wk after optic nerve crush. DD did not suppress axon regeneration. Optic nerve images were collected on a Zeiss LSM 710 laser scanning confocal microscope with a 10× objective, and tiled together by using the automatic stitching function of ZEN 2009. The images with the CTB channel were converted to grayscale and exported. (Scale bars: A and C, 50 μm; G, 100 μm.)