Figure S5.
Kif1C-5C chimera interferes with retrograde traffic (related to Fig. 7).(A) Cadherin-2 reporter particles appear larger and the background increased in THNs overexpressing the Kif1C-5C chimera. Yellow dotted lines indicate the area displayed in the line-scan analysis below the images. Scale bar, 10 µm. See also Video 6. (B) Background fluorescence level relative to the brightest Cadherin-2 reporter particle detected in the line-scan analyses suggest that the expression of Kif1C-5C interferes with endocytosis of the marker. n = 14 embryos/31 values for injection control, n = 13 embryos/25 tracks for p50-expressing embryos, n = 8 embryos/21 values for Kif5C-expressing embryos, 18 embryos/n = 30 values Kif5C T94N-expressing embryos, n = 9 embryos/19 values for Dynasore, and n = 22 values for Kif1C-5C–expressing embryos. Kruskal–Wallis ANOVA: P = 0.004 control/Kif5C T94N, P = 4.7 × 10−3 control/Dynasore, P = 6.4 × 10−8 control/p50, P = 6.10−8 Kif1C-5C/p50, and P = 5.10−4 Kif1C-5C/Kif5C T94N. (C) At the same time, large particles appear in chimera-expressing THNs. n = 14 embryos/164 values for injection control, n = 13 embryos/95 tracks for p50-expressing embryos, n = 8 embryos/124 values for Kif5C-expressing embryos, 18 embryos/n = 121 values Kif5C T94N-expressing embryos, n = 9 embryos/64 values for Dynasore, n = 114 values for Kif1C-5C–expressing embryos. Kruskal–Wallis ANOVA: P = 5.8 × 10−6 control/Kif1C-5C, P = 0.008 Kif5C T94N/Kif1C-5C, P = 0.002 control/Dynasore, P = 7.10−5 for Kif1C-5C/p50, P = 5.10−4 Kif1C-5C/Kif5C T94N. (D) Kif1C-5C reduces retrograde mean speeds of Cadherin-2 reporter particles more strongly than anterogradely moving particles. n = 593 tracks for Kif5C-expressing embryos and n = 439 tracks for Kif1C-5C–expressing embryos. (E) Similarly, maximal velocities in the plus direction are unaffected by chimera expression, while minus-directed velocities are strongly reduced compared with control. For full statistical testing, refer to Table S5 and Table S6. Anterograde: n = 399 tracks for injection control, n = 295 tracks for p50-expressing embryos, n = 579 tracks for Kif5C-expressing embryos, n = 425 tracks Kif5C T94N-expressing embryos, and n = 431 tracks for Kif1C-5C–expressing embryos. Retrograde: Anterograde: n = 411 tracks for injection control, n = 299 tracks for p50-expressing embryos, n = 580 tracks for Kif5C-expressing embryos, n = 432 tracks Kif5C T94N-expressing embryos, and n = 433 tracks for Kif1C-5C–expressing embryos. MHB, solid line; URL, dotted line. Boxes in graphs represent 25–75% of all values and whiskers 1.5 times the quartile. Median is shown as a horizontal bar and mean as a square box. Significance level in Kruskal–Wallis ANOVA: *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. All data shown in D and E represent n = 13 embryos for injection control, n = 14 p50-expressing embryos, n = 11 Kif5C-expressing embryos, n = 16 Kif5C T94N-expressing embryos, n = 8 embryos for Dynasore, and n = 9 Kif5C-1C–expressing embryos.

Kif1C-5C chimera interferes with retrograde traffic (related to Fig. 7 ). (A) Cadherin-2 reporter particles appear larger and the background increased in THNs overexpressing the Kif1C-5C chimera. Yellow dotted lines indicate the area displayed in the line-scan analysis below the images. Scale bar, 10 µm. See also Video 6. (B) Background fluorescence level relative to the brightest Cadherin-2 reporter particle detected in the line-scan analyses suggest that the expression of Kif1C-5C interferes with endocytosis of the marker. n = 14 embryos/31 values for injection control, n = 13 embryos/25 tracks for p50-expressing embryos, n = 8 embryos/21 values for Kif5C-expressing embryos, 18 embryos/n = 30 values Kif5C T94N-expressing embryos, n = 9 embryos/19 values for Dynasore, and n = 22 values for Kif1C-5C–expressing embryos. Kruskal–Wallis ANOVA: P = 0.004 control/Kif5C T94N, P = 4.7 × 10−3 control/Dynasore, P = 6.4 × 10−8 control/p50, P = 6.10−8 Kif1C-5C/p50, and P = 5.10−4 Kif1C-5C/Kif5C T94N. (C) At the same time, large particles appear in chimera-expressing THNs. n = 14 embryos/164 values for injection control, n = 13 embryos/95 tracks for p50-expressing embryos, n = 8 embryos/124 values for Kif5C-expressing embryos, 18 embryos/n = 121 values Kif5C T94N-expressing embryos, n = 9 embryos/64 values for Dynasore, n = 114 values for Kif1C-5C–expressing embryos. Kruskal–Wallis ANOVA: P = 5.8 × 10−6 control/Kif1C-5C, P = 0.008 Kif5C T94N/Kif1C-5C, P = 0.002 control/Dynasore, P = 7.10−5 for Kif1C-5C/p50, P = 5.10−4 Kif1C-5C/Kif5C T94N. (D) Kif1C-5C reduces retrograde mean speeds of Cadherin-2 reporter particles more strongly than anterogradely moving particles. n = 593 tracks for Kif5C-expressing embryos and n = 439 tracks for Kif1C-5C–expressing embryos. (E) Similarly, maximal velocities in the plus direction are unaffected by chimera expression, while minus-directed velocities are strongly reduced compared with control. For full statistical testing, refer to Table S5 and Table S6. Anterograde: n = 399 tracks for injection control, n = 295 tracks for p50-expressing embryos, n = 579 tracks for Kif5C-expressing embryos, n = 425 tracks Kif5C T94N-expressing embryos, and n = 431 tracks for Kif1C-5C–expressing embryos. Retrograde: Anterograde: n = 411 tracks for injection control, n = 299 tracks for p50-expressing embryos, n = 580 tracks for Kif5C-expressing embryos, n = 432 tracks Kif5C T94N-expressing embryos, and n = 433 tracks for Kif1C-5C–expressing embryos. MHB, solid line; URL, dotted line. Boxes in graphs represent 25–75% of all values and whiskers 1.5 times the quartile. Median is shown as a horizontal bar and mean as a square box. Significance level in Kruskal–Wallis ANOVA: *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. All data shown in D and E represent n = 13 embryos for injection control, n = 14 p50-expressing embryos, n = 11 Kif5C-expressing embryos, n = 16 Kif5C T94N-expressing embryos, n = 8 embryos for Dynasore, and n = 9 Kif5C-1C–expressing embryos.

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