Figure 2.
Identification of the minimum KIF1A domain required for cargo interaction. Overview of different KIF1Atd fragments, their localization, and interactions with cargo. (A, E, and I) Schematic depiction of the structure of truncated KIF1Atd constructs. (B, F, and J) Representative images of hippocampal neurons expressing KIF1Atd truncations (depicted in A, E, and I, respectively). (C, G, and K) Quantification of the polarity index of KIF1Atd truncations (depicted in A, E, and I, respectively). Dotted gray line marks the polarity index of KIF1Atd (N = 3, n = 32–40). (D, H, and L) Quantification of the normalized ratio of cargo intensity in axonal tips to that of the soma of hippocampal neurons coexpressing FRB-3myc–fused truncated KIF1Atd constructs (depicted in A, E, and I, respectively), marker-GFP, and FKBP-mRFP-KIF5Cmd, without or with addition of 1 µM rapalog. Dotted gray line marks the normalized intensity for KIF1Atd with rapalog treatment. Dotted red line marks the baseline of 5% of the KIF1Atd signal (N = 1, n = 7–15). (M) Overview of the identified interactions between truncated KIF1Atd and cargo. (N) Schematic depiction showing the interaction between KIF1A CC3 and PH domain with DCVs, lysosomes, and SVs. Data are displayed as mean ± SEM. Mann–Whitney U test; ***, P < 0.001. Scale bars, 20 µm.

Identification of the minimum KIF1A domain required for cargo interaction. Overview of different KIF1Atd fragments, their localization, and interactions with cargo. (A, E, and I) Schematic depiction of the structure of truncated KIF1Atd constructs. (B, F, and J) Representative images of hippocampal neurons expressing KIF1Atd truncations (depicted in A, E, and I, respectively). (C, G, and K) Quantification of the polarity index of KIF1Atd truncations (depicted in A, E, and I, respectively). Dotted gray line marks the polarity index of KIF1Atd (N = 3, n = 32–40). (D, H, and L) Quantification of the normalized ratio of cargo intensity in axonal tips to that of the soma of hippocampal neurons coexpressing FRB-3myc–fused truncated KIF1Atd constructs (depicted in A, E, and I, respectively), marker-GFP, and FKBP-mRFP-KIF5Cmd, without or with addition of 1 µM rapalog. Dotted gray line marks the normalized intensity for KIF1Atd with rapalog treatment. Dotted red line marks the baseline of 5% of the KIF1Atd signal (N = 1, n = 7–15). (M) Overview of the identified interactions between truncated KIF1Atd and cargo. (N) Schematic depiction showing the interaction between KIF1A CC3 and PH domain with DCVs, lysosomes, and SVs. Data are displayed as mean ± SEM. Mann–Whitney U test; ***, P < 0.001. Scale bars, 20 µm.

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