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. 2018 Jul 17;24(3):685–700. doi: 10.1016/j.celrep.2018.06.071

Figure 5.

Figure 5

DCV Dendritic Trafficking Is KIF1A Dependent and TANC2/Liprin-α Independent

(A) Neurons (11–14 DIV) co-transfected with GFP-KIF1A (green), in combination with NPY-RFP, mCherry-Syt4, or mCherry-Syt11 (red). Arrows point to co-localizing puncta. Scale bar, 5 μm. See also Figures S3A and S3B.

(B and C) Percentage of co-localizing puncta between DCV proteins and KIF1A (B), and vice versa (C). The bars show mean ± SEM (n = 19–23 dendrites).

(D) NPY-GFP (green), in combination with mCherry-Syt4 or mCherry-Syt11 (red). Scale bar, 5 μm. See also Figures S3C and S3D.

(E and F) Co-localizing puncta between NPY and Syt4 or Syt11 (E), and vice versa (F). The bars show mean ± SEM (n = 21–31 dendrites).

(G) Phluorin-Syt4 (green) with mCherry-Syt11 (red) or Phluorin-Syt11 (green) with mCherry-Syt4 (red). Arrows point to co-localizing puncta. Scale bar, 5 μm. See also Figures S3E and S3F.

(H and I) Co-localizing puncta between Syt4 and Syt11 (H), and vice versa (I). The bars show mean ± SEM (n = 20–27 dendrites).

(J and K) Representative kymographs showing trajectories of NPY-RFP vesicles (J) or mCherry-Syt4 vesicles (K) (red) moving with GFP-KIF1A (green).

(L) Representative images acquired at 10 s intervals showing NPY-GFP motility in dendrites of neurons transfected with pSuper (right) or shRNA_KIF1A (left). In the merged images, red corresponds to 0 s, blue corresponds to 10 s, and green corresponds to 20 s. Scale bar, 5 μm.

(M) Quantification of the percentage of mobile NPY vesicles in neurons (11–14 DIV) co-transfected with NPY-GFP, in combination with pSuper, shRNA_KIF1A, shRNA_KIF1A + FL-HA-KIF1A, or shRNA_KIF1A + HA-KIF1A(1-1105). The bars show mean (n = 18–32 dendrites; ∗p < 0.05, ∗∗p < 0.001, ∗∗∗p < 0.001, t test).

(N and O) Quantification of the percentage of mobile DCVs in neurons (11–14 DIV) co-transfected with NPY-GFP (N) or mCherry-Syt4 (O), in combination with pSuper, shRNA_KIF1A, shRNA_TANC2, shRNA_liprin-α2-3, or shRNA_liprin-α2-3 + shRNA_TANC2. The bars show mean (n = 17–58 dendrites in N and n = 17–39 dendrites in O; ∗∗∗p < 0.001, t test). See also Figures S3J–S3L.