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. 2014 Dec 15;127(24):5179–5188. doi: 10.1242/jcs.149633

Fig. 4.

Fig. 4.

KIF1C associates with CLASP-rich MTs. (A,B) CLASPs (green, white arrows) mark MT plus ends in a non-treated cell (A), but upon PDBu treatment (B), CLASPs accumulate at the lattice of MTs in close contact with podosomes (cortactin, red). Data show wide-field fluorescence analysis of immunostaining. (C) KIF1C–mCherry (red, yellow arrows) moves with the end of a GFP–CLASP2-associated MT (green, white arrows) in a PDBu-treated cell. Data show frames from a video sequence and are enlarged from the box in supplementary material Fig. S1F. See also supplementary material Movies 5, 6. (D) KIF1C (red) relocation events occurring at the ends of MTs associated with CLASP2 (CL-MT), as compared to the same events superimposed on a spatially shifted RFP–CLASP2 video sequence, based on data as in supplementary material Movies 5, 6. Data show the mean+s.e.m. (N = 4 cells, 14–19 events/cell); *P<0.001 (Student's t-test). (E,F) Colocalization of ectopically expressed KIF1C with mitochondria in cells with (mitoCL) or without (ctr) mito-CLASP expression. Mander's coefficients M1 (E) and M2 (F) for thresholded images are shown. Both full-length (left) and (right) and tail domain constructs are recruited to mitochondria by CLASP. Data show the mean+s.e.m. (N = 5); *P<0.01 (Student's t-test). (G,H) KIF1C–GFP (red) colocalizes with CLASP (green, immunostained) and mitochondria (mCherry–TOM20, blue) in cells with (H) but not without (w/o, G) mito-CLASP. (I,J) FLAG-tagged KIF1C tail (red) colocalizes with CLASP (green, immunostained) and Mitotracker (blue) in cells with (J) but not without (I) mito-CLASP. See also supplementary material Fig. S2A–D. (K,L) Mitochondria in cells transfected with mCherry–TOM20 alone (K) or mCherry–TOM20 and mito-CLASP (L). See also supplementary material Fig. S2F,G. Yellow dotted lines indicate cell borders. (M) The number of mitochondria in lamellae in cells transfected with mCherry–TOM20 or mito-CLASP, based on the data as shown in K,L. Data show the mean+s.e.m. (N = 28–36); *P<0.01 (Student's unpaired two-tailed t-test).