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. 2021 May 20;220(7):e202010179. doi: 10.1083/jcb.202010179

Figure 1.

Figure 1.

Multiple dynein effectors associate with axonal autophagosomes. (A–C) Time series and kymographs from separate LC3+ puncta demonstrating comigration with JIP1, HAP1, and JIP3. Scale bar, 2 µm. (D) Schematic illustrating neuronal subregions and axonal transport of autophagosomes. (E–G) Quantification of LC3+ puncta comigrating with JIP1, HAP1, and JIP3 in different axonal regions. n = 9–17 neurons; one-way ANOVA with Tukey’s multiple comparisons test (JIP1: distal or mid vs. proximal, P < 0.0001). (H and I) Immunoblotting and quantification (relative to brain lysate, input) of autophagosome isolation illustrating enrichment on the outer membrane. n = 3–4 preps; one-way ANOVA (LC3, P < 0.0001; LIC1, P = 0.0056; JIP1, P = 0.0497; HAP1, P = 0.0058; JIP3, P = 0.0277). (J and K) Micrographs and quantification showing PLA puncta for endogenous DIC with Halo-JIP1 along the axon (dotted gray line). Arrowheads indicate PLA puncta. Scale bar, 10 µm. Dashed gray line indicates negative control (missing primary antibody). n = 7 neurons; one-way ANOVA with Tukey’s multiple comparisons test (distal vs. mid, P = 0.0236; distal vs. proximal, P = 0.0131). Bars throughout show mean ± SEM. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.