Fig. 2: KIF11 regulates EB3 dynamics in mature neurons. | Nature Communications

Fig. 2: KIF11 regulates EB3 dynamics in mature neurons.

From: Intellectual disability-causing mutations in KIF11 impair microtubule dynamics and dendritic arborization

Fig. 2: KIF11 regulates EB3 dynamics in mature neurons.The alternative text for this image may have been generated using AI.

A Experimental timeline. B Left: Maximum projection intensity images of an axon from cells co-expressing NC-GFP (scrambled negative control) and miRFP703-EB3 (far-red tagged EB3 protein). Red arrows (B–E) point to the base of the selected projection. Right: Selected axon and kymograph of miRFP703-EB3. For all kymographs (B–E), the vertical arrow represents distance, with the base of the arrow positioned towards the soma and the arrowhead positioned towards the tip of the projection. The horizontal arrows represent time progressing from left to right. C Left: Maximum projection intensity images of axons from cells co-expressing shKif11 (shRNA targeting KIF11) and miRFP703-EB3. Right: Selected axon and kymograph of miRFP703-EB3. D Left: Maximum projection intensity images of dendrites from cells co-expressing NC-GFP (scrambled negative control) and miRFP703-EB3. Right: Selected dendrite and kymograph of miRFP703-EB3. E Left: Maximum projection intensity images of dendrites from cells co-expressing shKif11 (shRNA targeting KIF11) and miRFP703-EB3. Right: Selected dendrite and kymograph of miRFP703-EB3.The percentage of minus-end-out MTs in axons (F) and dendrites (G) in NC-GFP or shKIF11 neurons. Two-tailed Unpaired t-test. EB3-comet flux in axons (H) and dendrites (I) in NC-GFP or shKIF11 neurons. One-way ANOVA, Tukey’s test. EB3-comet growth rate in axons (J) and dendrites (K) in NC-GFP or shKIF11 neurons. One-way ANOVA, Tukey’s test. EB3-comet distance traveled (MT growth) in axons (L) and dendrites (M) in NC-GFP or shKIF11 neurons. One-way ANOVA, Tukey’s multiple comparison test. N The percentage of minus-end-out MTs in primary, secondary, and tertiary dendrites. Mixed-effects model (REML) followed by Tukey’s test. O EB3-comet flux for plus-end-out and minus-end-out EB3 comets in secondary dendrites. One-way ANOVA, Tukey’s test. For all graphs (F–O), error bars represent ±SEM. P-values are listed above respective comparisons. For F, H, J, L, N = 10, 16 neurons and axons for NC-GFP or shKIF11, respectively. For G, I, K, M, N = 16(40), 15(29) neurons (dendrites) for NC-GFP and shKIF11, respectively. For O and N, NC-GFP N = 16 neurons (4 primary, 14 secondary, and 14 tertiary dendrites), shKIF11 N = 15 neurons (4 primary, 16 secondary, and 9 tertiary dendrites). Source data are provided as a Source Data file.

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