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. 2014 May 27;55(5):3237–3246. doi: 10.1167/iovs.13-13336

Figure 5.

Figure 5

Autophagy activation in the retinas of Gnat1−/− and Sag−/− mouse strains. (A–C) Western blots of LC3 and quantification of LC3-I to LC3-II ratios in Group 1, Group 2, and Group 3 Gnat1−/− animals, respectively, as described in Methods. There is loss of the light-period peak of LC3-II and Atg5-Atg12 complex formation in Group 1 animals, with the ratio at 10 PM being significantly higher than that at 1 PM (P < 0.05, n = 8 animals per time point). Note that the dark-period peak occurs earlier in the Gnat1−/− mice as compared to wild-type controls. There is no loss of the dark-period peak, either during regular 12-hour light/12-hour dark cycle (A) or in Group 2 animals shifted from light back to dark at 9 AM, or after 3 hours in the light (B). (D–F) Western blots and quantification of LC3-I to LC3-II ratios in Group 1, Group 2, and Group 3 Sag−/− mice, respectively. There is loss of the dark-period peak of LC3-II and Atg5-Atg12 complex formation in Group 1 animals, with preservation of the light-period peak (P < 0.05, n = 8 animals per time point). Group 2 animals shifted from light back to dark at 9 AM, after 3 hours in the light, show a decrease in LC3-II levels, but no time-dependent regeneration of this isoform (E). Maintaining the animals in the dark resulted in a gradual decrease in LC3-II levels.