Fig. 3: VPS35 deficiency dysregulates OS morphology, OS protein turnover, and endomembrane homeostasis in rods.

a, b Low (left) and high (right) magnifications of electron micrographs showed the OS in 1-month-old Ctrl and KO mice. tv: tubulovesicles. c Immunoblots of indicated OS proteins. As expected, rhodopsin has a propensity to form monomers, dimers, and trimers. Bar graphs show the relative intensity of protein bands of indicated molecules (normalized using α-tubulin as an input control) considering Ctrl as 1. d 3-month-old mouse retinas stained by indicated markers. Arrows mark the residual EEA1 signals and increased CD63-labeled LEs at the IS-ONL borders. Arrowheads mark the increased OPL expression of CD63-labeled LEs. Insets: (Top) Electron micrograph shows Ctrl rod IS has solitary EE globes surrounded by tubules (t). (Bottom) EE clusters are collapsed in the lower part of the KO rod IS. Bars in nm. e, f Bar graphs show the fold change in signal density (counts per 1,1000 mm²) of indicated markers in IS (e) and OPL (f). For EEA1, N = 3 mice each for both Ctrl and KO (1 image per animal); for CD63, N = 3 mice for Ctrl and N = 4 mice for KO animals (1 image per animal); for Lamp1 and LC3, N = 3 mice each for both Ctrl and KO (two images per animal) are shown. g LC3 immunoblots of in 3-month-old mouse retinas and intensity of the LC3I and LC3II signal (normalized using α-tubulin as an input control) considering Ctrl as 1. h Co-staining of Lamp1 and LC3 in the indicated retinal layers of 3-month-olds. Arrows mark LC3-postive, Lamp1-negative/ puncta. For c, e–g, Mean ± SD and P-values of N = 3 in each group are shown. Two-tailed Student’s t-test. Representative images of 3 independent repeats (a–e, g, h) are shown. Source data of (c, e–g) are provided as a Source Data file.