Picture from Niwa S et al. (2017) Curr Biol "BORC Regulates the Axonal Transport of Synaptic Vesicle Precursors by ...."

Figure S7 KXD-1 co-localizes with SVs marked by SNB-1 (related to Figure 7)On the left, confocal images of SNB-1::YFP (green) and KXD-1::mRuby (magenta) in straightened DA9 dorsal axons,which are aligned at the commissure bend on the right, of three wild-type animals visualized using wyIs92 andwyEx9053. Scale bar represents 5 um. On the right, corresponding DA9 cell bodies from the same animals. Scale bar represents 2.5 um.

Picture from Niwa S et al. (2017) Curr Biol "BORC Regulates the Axonal Transport of Synaptic Vesicle Precursors by ...."

(A) Representative ARL-8::YFP and mCherry::RAB-3kymographs in the ventral axon of WT andsam-4(tm3828) animals (wyIs546). Each dotted linerepresents one event; co-movement is visualizedas staggering between dotted lines of differentcolors. ARL-8 co-migrates with SVPs marked bymCherry::RAB-3 in WT but is detached from SVPsand largely diffused in sam-4(tm3828). Scale barrepresents 2.5 um.(B) Quantification of the percentage of ARL-8-positive anterograde RAB-3 events in WT and sam4(tm3828) animals. The percentage is calculated asthe number of ARL-8-positive anterograde RAB-3events over the total number of anterograde RAB-3events. Lines denote median values, and each dotrepresents one animal. Mann-Whitney test, ****twotailed p value < 0.0001; n = 24 and 25 animals for WTand sam-4(tm3828), respectively.

Picture from Niwa S et al. (2017) Curr Biol "BORC Regulates the Axonal Transport of Synaptic Vesicle Precursors by ...."

Figure S1 RAB-7 and LAAT-1 show distinct localization patterns from RAB-3 (related to Figure 1)(A) On the left, confocal images of GFP::RAB-7 (late endosome marker) in straightened DA9 dorsal axons,which are aligned at the commissure bend on the right and stacked in rows with one axon in each row, ofthree wild-type animals visualized using wyEx4837. Scale bar represents 5 um. On the right, corresponding DA9 cell bodies from the same animals. Scale bar represents 2.5 um.(B) On the left, confocal images of LAAT-1::GFP (lysosome marker) in straightened DA9 dorsal axons, whichare aligned at the commissure bend on the right and stacked in rows with one axon in each row, of threewild-type animals visualized using wyEx8985. Scale bar represents 5 um. On the right, correspondingDA9 cell bodies from the same animals. Scale bar represents 2.5 um.

Picture from Niwa S et al. (2017) Curr Biol "BORC Regulates the Axonal Transport of Synaptic Vesicle Precursors by ...."

(A) A schematic drawing of the DA9 neuron. The dorsal asynaptic region, the ventral axon, and the commissure that are observed and analyzed in this study areshown. Asterisk indicates the location where the commissure joins the dorsal nerve cord.(B-F) Representative images of GFP::RAB-3 in wild-type (WT) (B), sam-4(tm3828)

Picture from Niwa S et al. (2017) Curr Biol "BORC Regulates the Axonal Transport of Synaptic Vesicle Precursors by ...."

Figure 1. SAM-4 and ARL-8 Are in the Same Pathway(A) A schematic drawing of the DA9 neuron. The dorsal asynaptic region, the ventral axon, and the commissure that are observed and analyzed in this study areshown. Asterisk indicates the location where the commissure joins the dorsal nerve cord.(B-F) Representative images of GFP::RAB-3 in wild-type (WT) (B), sam-4(tm3828) (C), arl-8(tm2388) (D), arl-8(wy271) (E), and arl-8(wy271); sam-4(tm3828) (F).GFP::RAB-3 was expressed using the itr-1 pB promoter. Asterisks indicate the commissure bend shown in (A). Scale bars, 50 um.(G) Image montages of the dorsal axons in WT, sam-4(3828), arl-8(tm2388), arl-8(wy271), and sam-4(tm3828); arl-8(wy271). 10 confocal images from eachgenotype were cropped, straightened, and aligned in rows with the commissure bend (asterisk) on the right using ImageJ. Scale bar, 20 um.(H and I) Statistical analysis of mutant phenotypes. The number of puncta mis-accumulated to the commissure (H) and the length of the asynaptic region (I) areshown. Lines show median values, and each dot represents one animal. Kruskal-Wallis one-way ANOVA on ranks and Dunn's multiple comparisons test,*adjusted p value < 0.05, **adjusted p value < 0.01, and ***adjusted p value < 0.001; n = 20 animals for each genotype. See also Figure S1.

Picture from Niwa S et al. (2017) Curr Biol "BORC Regulates the Axonal Transport of Synaptic Vesicle Precursors by ...."

Figure 6. BLOS-9 Regulates the AxonalTransport of SVs Together with SAM-4(A-F) Image montages of the dorsal axons in WT (A),blos-9(jpn2) (B), blos-9(jpn2) expressing BLOS-9using the DA9 promoter (C), blos-9(jpn2); sam4(tm3828) (D), blos-9(jpn2); arl-8(jpn1) (E), and blos9(jpn2); unc-104(wy873) (F). Note that arl-8(jpn1)and unc-104(wy873) are gain-of-function alleles.Scale bar, 20 um.(G and H) Statistical analysis of suppressormutants. The number of GFP::RAB-3 puncta misaccumulated to the commissure (G) and the lengthof the asynaptic region (H) are shown. Lines showmedian values, and each dot represents oneanimal. Kruskal-Wallis one-way ANOVA on ranksand Dunn's multiple comparisons test, *adjustedp value < 0.05, **adjusted p value < 0.01, and***adjusted p value < 0.001; n = 20 animals for eachgenotype.(I-L) The localization of BLOS-9::GFP (I) andmCherry::RAB-3 (J) in the dorsal synaptic region.The merged image is shown in (K). (L) is a zoomedin image of the boxed area in (K). BLOS-9 andRAB-3 are well co-localized. Scale bars, 50 um.See also Figure S5.

Picture from Niwa S et al. (2017) Curr Biol "BORC Regulates the Axonal Transport of Synaptic Vesicle Precursors by ...."

Figure 3. ARL-8 Dynamics in the sam-4Mutant(A) Representative ARL-8::YFP and mCherry::RAB-3kymographs in the ventral axon of WT andsam-4(tm3828) animals (wyIs546). Each dotted linerepresents one event; co-movement is visualizedas staggering between dotted lines of differentcolors. ARL-8 co-migrates with SVPs marked bymCherry::RAB-3 in WT but is detached from SVPsand largely diffused in sam-4(tm3828). Scale barrepresents 2.5 um.(B) Quantification of the percentage of ARL-8-positive anterograde RAB-3 events in WT and sam4(tm3828) animals. The percentage is calculated asthe number of ARL-8-positive anterograde RAB-3events over the total number of anterograde RAB-3events. Lines denote median values, and each dotrepresents one animal. Mann-Whitney test, ****twotailed p value < 0.0001; n = 24 and 25 animals for WTand sam-4(tm3828), respectively.(C) Quantification of the number of moving vesiclesin the proximal asynaptic region of axon, normalizedto the duration and the length of the axon (n = 20animals; t test, ***p < 0.001, **p < 0.01, and*p < 0.05). Error bars represent SEM.(D) Diagram and kymograph showing a dissociationevent (arrow) and quantification of the dissociationrate at stable puncta in the proximal asynaptic regionof axon (n = 20 animals; t test, ***p < 0.001 and**p < 0.01). Error bars represent SEM.(E) Diagram and kymograph showing a mobilepacket captured by a stable punctum (arrow) andquantification of the capture probability in the ventralaxon (n = 20 stable puncta; t test, no statisticalsignificance). Error bars represent SEM.See also Figure S3.

Picture from Niwa S et al. (2017) Curr Biol "BORC Regulates the Axonal Transport of Synaptic Vesicle Precursors by ...."

(I-L) The localization of BLOS-9::GFP (I) andmCherry::RAB-3 (J) in the dorsal synaptic region.The merged image is shown in (K). (L) is a zoomedin image of the boxed area in (K). BLOS-9 andRAB-3 are well co-localized. Scale bars, 50 um.See also Figure S5.

Picture from Meng L et al. (2016) PLoS Genet "Regulation of Gap Junction Dynamics by UNC-44/ankyrin and UNC-33/CRMP through ...."

Representative images of immunostaining results for FLAG::UNC-33(S)(A) and UNC-44(B) in control and mutant animals. UNC-33(S) localization was determined by a transgene expressing FLAG tagged UNC-33(S) in all neurons (Prgef-1::FALG::unc-33(S)).

Picture from Meng L et al. (2016) PLoS Genet "Regulation of Gap Junction Dynamics by UNC-44/ankyrin and UNC-33/CRMP through ...."

Representative images of immunostaining results for FLAG::UNC-33(S)(A) and UNC-44(B) in control and mutant animals. UNC-33(S) localization was determined by a transgene expressing FLAG tagged UNC-33(S) in all neurons (Prgef-1::FALG::unc-33(S)).