LE-loaded dynein–snapin complexes drive amphisome retrograde trafficking. (A and B) Kymographs (A) and quantitative analysis (B) showing impaired retrograde transport of amphisomes by disrupting DIC–snapin coupling. DRG neurons were cotransfected with GFP-LC3 and mRFP-Rab7 along with HA-snapin, HA-snapin-L99K, or HA vector at DIV0 and time-lapse imaged for 3 min at DIV3. The total number of neurons (N) examined is indicated in parentheses from greater than three experiments. (C and D) Images (C) and quantitative analysis (D) showing that disrupted snapin–DIC coupling increases the density of axonal amphisomes. The total number of neurons examined for each group is 30 from more than three experiments. (E) Disrupting snapin–DIC coupling had no significant effect on fusion between autophagosomes and LEs. Data were quantified from total number of AVs denoted in or above the bars from greater than three experiments. Mann–Whitney test (B) and Student’s t test (D and E). Error bars: SEM. Bars: (A) 10 µm; (C) 5 µm.