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. 2018 Jul 18;7:1107. [Version 1] doi: 10.12688/f1000research.15409.1

Figure 8. Intracellular trafficking and processing of photoreceptor outer segments in a feeding assay.

Figure 8.

Cultures were pulsed with photoreceptor outer segments (POS) bound to FITC. Cargo internalisation and trafficking via phagosomes/endosomes, lysosomes and autophagy bodies were quantified at 2, 4, 6, 12, 24 and 48 hours. Following internalisation, POS were detected [ A– B] in Rab 5 early vesicles by 2–4 hours. These diminished over time as cargo appeared [ C– D] in Rab 7-positive compartments. [ E– F] At 6 hours, a large proportion of POS were present in early lysosomes and [ G– H] in LAMP2 vesicles by 12–24 hours. [ I– J] Cargos appeared in LC3B-positive autophagy bodies afterwards which were present up to 48 hours. Images show representative confocal images with quantification (n=10 cells/compartment/time point) in 2 independent experiments. Inserts show extent of co-localisation in red (R) and green (G) channels using an unbiased quantification method by Volocity. Data is presented as mean ± SEM and sourced in part from material published previously 18, 50 under the Creative Commons licence.