All animal studies adhered to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and were approved by the institutional animal care and use committee at Baylor College of Medicine. Wild type (WT) C57BL/6J (#000664), B6 albino (#000058 B6[Cg])-
Tyrc-2J/J), and Pcp2-Cre (#006207 L7 Cre, Tg[Pcp2-cre] 1Amc/J) mice were purchased from the Jackson Laboratory (Bar Harbor, ME, USA). The Pcp2-Cre mice were found to have the Crb1
rd8 allele
20 and were crossed with WT C57BL6/J for at least five generations. Although this Pcp2-Cre line was reported by the vendor (Jackson Laboratory) also to express green fluorescent protein (GFP), in retinal bipolar cells or in Purkinje cells in which we could clearly observe Cre immunostaining (
Supplementary Fig. S1), we did not observe GFP fluorescence under our imaging conditions using our immunostaining protocol (
Supplementary Fig. S2). Vps34 floxed mice (Vps34
fl/fl), which have loxP sites flanking exons 17 and 18,
13 were a gift from Fan Wang (Duke University); these mixed background C57BL6/129 mice were back-crossed with WT C57BL/6J for at least six generations. Vps34 floxed mice then were bred with Pcp2-Cre mice to generate conditional Vps34 KO mice (Vps34
fl/fl;Pcp2-Cre). The KO mice suffered from progressive ataxia and generally were euthanized for animal welfare reasons when they developed difficulty in feeding, which usually happened before 10 months of age. Mice with a GFP-LC3 transgene
21 (#RBRC00806, GFP-LC3#53) were obtained from the RIKEN Bio Resource Center (Ibaraki, Japan). Vps34 KO mice also containing transgenic GFP-LC3 (Vps34
fl/fl;Pcp2-Cre;GFP-LC3
+/-) were generated by crossing Vps34
fl/fl;Pcp2-Cre mice with the GFP-LC3 transgenic mice. Only mice heterozygous for the GFP-LC3 allele were used.