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[Preprint]. 2023 Feb 26:2023.02.26.530068. [Version 1] doi: 10.1101/2023.02.26.530068

Figure 2: LRK-1 and AP-3 act in parallel and through SYD-2 to regulate lysosomal protein trafficking.

Figure 2:

(A) Quantitation of co-transport of SNG-1::eGFP and mCherry::RAB-3 in WT, lrk-1(km17), and apb-3(ok429) from kymograph analysis of sequential dual color imaging at 1.3 frames per second (fps). # P-value ≤ 0.05 (One-Way ANOVA with Tukey’s post-hoc test, all comparisons to WT); ns: not significant; Number of animals (N) ≥ 15 per genotype; Number of vesicles analyzed per genotype (n) > 800.

(B) Quantitation of fraction of SNG-1-carrying vesicles co-transporting CTNS-1 in WT, lrk-1(km17), lrk-1(km41), apb-3(ok429) and lrk-1(km17) apb-3(ok429), from kymograph analysis of sequential dual color imaging at 1.3 fps. # P-value ≤ 0.05 (Mann–Whitney Test); N ≥ 15 per genotype; n > 500.

(C) Quantitation of fraction of SNG-1-carrying vesicles co-transporting RAB-7 from WT, lrk-1(km17), apb-3(ok429) and lrk-1(km17) apb-3(ok429), kymograph analysis of dual color imaging. # P-value ≤ 0.05 (One-Way ANOVA with Tukey’s post-hoc test, all comparisons to WT); ns: not significant; N ≥ 20 per genotype; n > 800.

(D) CTNS-1::mCherry in the cell body, process, and synapses of PLM neurons of WT, lrk-1(km17), apb-3(ok429), and lrk-1(km17) apb-3(ok429). Scale bar: 10 μm. Red arrows point to some CTNS-1-labelled compartments.

(E) mScarlet::RAB-7 in the cell body, process, and synapses of PLM neurons of WT, lrk-1(km17), apb-3(ok429), and lrk-1(km17) apb-3(ok429). Scale bar: 10 μm. Red arrows point to some RAB-7-labelled compartments.

(F) LMP-1::mNeonGreen in the cell body, process, and synapses of PLM neurons of WT, lrk-1(km17), apb-3(ok429), and lrk-1(km17) apb-3(ok429). Scale bar: 10 μm. Red arrows point to some RAB-7-labelled compartments.

(G) Images showing APB-3::GFP puncta in the head ganglion cell bodies of WT, lrk-1(km17), and syd-2(ok217). Scale bar: 10 μm. Red boxes highlight the regions of insets with cell bodies from images showing APB-3::GFP in (i) WT, (ii) lrk-1, and (iii) syd-2.

(H) Quantitation of the number of APB-3::GFP puncta per cell body in WT, lrk-1(km17), and syd-2(ok217). # P-value ≤ 0.05 (Mann–Whitney Test); ns: not significant; N > 10 animals; n > 75 cell bodies.

(I) Quantitation of intensity of APB-3::GFP puncta in cell bodies of WT, lrk-1(km17), and syd-2(ok217). The ratio of the intensity of APB-3::GFP puncta to cytosolic intensity in the cell body is plotted. # P-value ≤ 0.05 (Mann–Whitney Test); ns: not significant; N > 10 animals; n > 75 cell bodies.

(J) Quantitation of average size of APB-3::GFP puncta per cell body in WT, lrk-1(km17), and syd-2(ok217). # P-value ≤ 0.05 (Mann–Whitney Test); ns: not significant; N > 10 animals; n > 75 cell bodies.