kif11 oe  (OriGene)

Journal: Nature Communications

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

doi: 10.1038/s41467-026-70522-z

Figure Lengend Snippet: A Schema of selected Microcephaly with or without chorioretinopathy, lymphedema, or intellectual disabilities (MCLID) patient mutations ( Hs: Homo sapiens ) and the corresponding mouse homolog (Mm: Mus Musculus ) (adapted from Schlögel et al. ) on the KIF11 protein. B Experimental timeline. C Confocal projection images of primary hippocampal mouse neurons transfected with control or KIF11 constructs, with the soma in the center of the image. Scale Bar=25 µm. D Soma size quantification of ( C ). N = 20,22,22,19 neurons for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Tukey’s test. E Quantification of dendritic morphology changes using Sholl analysis. N = 17,20,20,16 neurons for NC-GFP, KIF11-OE, KIF11 Y81F, and KIF11 ΔCterm , respectively. Two-way ANOVA followed by Tukey’s test. F Plus-end-out EB3-comet flux in KIF11 dendrites in comparison to NC-GFP. N = 17,17,18,21 dendrites for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. G . Minus-end-out EB3-comets flux in KIF11 dendrites in comparison to NC-GFP. N = 14,16,18,21 dendrites for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. H Percentage of Minus-end-out EB3-comets in KIF11 dendrites in comparison to NC-GFP. N = 17,17,18,22 dendrites for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. I . Length of plus-end-out MT growth for KIF11 dendrites in comparison to NC-GFP. N = 17(248),17(59),18(114),22(184) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. J Length of minus-end-out MT growth for KIF11 dendrites compared to NC-GFP. N = 17(70),17(42),18(27),22(45) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. K Plus-end-out MT growth-rate based on EB3-comet velocities in KIF11 dendrites in comparison to NC-GFP. N = 17(254),17(56),18(130),22(189) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. L Minus-end-out MT growth rate based on EB3-comet velocities in KIF11 dendrites in comparison to NC-GFP. N = 17(59),17(49),18(32),22(41) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. For all graphs ( D–L ), error bars represent ±SEM. P values are listed above respective comparisons. P values are listed above respective comparisons. Source data are provided as a file.

Article Snippet: DIV16 primary hippocampal neurons culture (in 35 mm Mattek dishes) were transfected with 0.5 μg of NC-GFP (Origene TR30013) or KIF11-OE (Genecoepia CS-Mm03465-Lv236-01) for Supplementary Fig. ; or 0.5 μg NC-GFP (Origene TR30013), KIF11-OE (Genecoepia CS-Mm03465-Lv236-01), or Kif11 Y81F (Genecoepia CS-Mm03465-Lv236-02) for Supplementary Fig. . At DIV18, neuronal cultures were washed with PBS and then fixed in a freshly prepared solution of 4% paraformaldehyde for 15 min. After three or more rinses in PBS, the cells were then incubated in 10% normal horse serum(NHS) (Gibco) in PBS-T with 0.1% Triton X-100 for 1 h at room temperature to reduce the non-specific binding of the primary antibody.

Techniques: Transfection, Control, Construct, Comparison

Journal: Nature Communications

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

doi: 10.1038/s41467-026-70522-z

Figure Lengend Snippet: A Timeline of experimental design to record miniature excitatory post-synaptic potential (mEPSCs) in mouse primary hippocampal culture expressing NC-GFP or KIF11 constructs. Tetrodotoxin (TTX) was added to ensure mEPSCs and not spontaneous EPSCs were captured. B Two representative traces of mEPSCs for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm 24–48 h post-transfection. Bar graph of mEPSC amplitude ( C ) and frequency ( D ) in NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm neurons. N = 10,13,12,13 NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm neurons respectively. One-way ANOVA followed by Tukey’s Multiple comparisons test. Cumulative probability graphs showing no change in mEPSC amplitude ( E ), but reduced frequency ( F ) in KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm neurons compared to NC-GFP. Kolmogorov-Smirnov Test. For graphs ( C , D ), error bars represent ± SEM. P-values are listed above respective comparisons. Source data are provided as a Source Data file.

Article Snippet: DIV16 primary hippocampal neurons culture (in 35 mm Mattek dishes) were transfected with 0.5 μg of NC-GFP (Origene TR30013) or KIF11-OE (Genecoepia CS-Mm03465-Lv236-01) for Supplementary Fig. ; or 0.5 μg NC-GFP (Origene TR30013), KIF11-OE (Genecoepia CS-Mm03465-Lv236-01), or Kif11 Y81F (Genecoepia CS-Mm03465-Lv236-02) for Supplementary Fig. . At DIV18, neuronal cultures were washed with PBS and then fixed in a freshly prepared solution of 4% paraformaldehyde for 15 min. After three or more rinses in PBS, the cells were then incubated in 10% normal horse serum(NHS) (Gibco) in PBS-T with 0.1% Triton X-100 for 1 h at room temperature to reduce the non-specific binding of the primary antibody.

Techniques: Expressing, Construct, Transfection

Journal: Nature Communications

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

doi: 10.1038/s41467-026-70522-z

Figure Lengend Snippet: A Experimental timeline for in vivo expression of MCLID mutations in mouse. In utero electroporation (IUE) was used to insert eGFP, WT KIF11 (KIF11-OE), KIF11 Y81F , or KIF11 ΔCterm constructs into the hippocampi of embryonic day 15.5 mice. These mice developed until postnatal day 7 or day 21. Their brains were extracted, fixed in 4% PFA, cryoprotected, coronally sectioned at 50 µm, and imaged on a confocal microscope. B Representative images of neurons expressing eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm in the dorsal CA1 of P7 mice. C Sholl analysis of neurons expressing eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm in the dorsal CA1 of P7 mice. Two-way ANOVA followed by Tukey’s multiple comparisons test. D Number of branch points per neuron in P7 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. E Individual branch lengths per neuron in P7 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. F Sum of branch lengths per neuron in P7 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. N = 10(3),13(3),13(2),13(3) neurons (Mice) in eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm P7 mice, respectively. G Representative images of neurons expressing eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm in the dorsal CA1 of P21 mice. H Sholl analysis of neurons expressing eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm in the dorsal CA1 of P21 mice. Two-way ANOVA followed by Tukey’s multiple comparisons test. I Number of branch points per neuron in P21 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. J Individual branch lengths per neuron in P21 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. K Sum of branch lengths per neuron in P21 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. N = 10(3),10(3),10(2), 10(3) neurons (Mice) in eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm P21 mice respectively. For all graphs ( D–F , I–K ), error bars represent ±SEM. P values are listed above respective comparisons. Source data are provided as a file.

Article Snippet: DIV16 primary hippocampal neurons culture (in 35 mm Mattek dishes) were transfected with 0.5 μg of NC-GFP (Origene TR30013) or KIF11-OE (Genecoepia CS-Mm03465-Lv236-01) for Supplementary Fig. ; or 0.5 μg NC-GFP (Origene TR30013), KIF11-OE (Genecoepia CS-Mm03465-Lv236-01), or Kif11 Y81F (Genecoepia CS-Mm03465-Lv236-02) for Supplementary Fig. . At DIV18, neuronal cultures were washed with PBS and then fixed in a freshly prepared solution of 4% paraformaldehyde for 15 min. After three or more rinses in PBS, the cells were then incubated in 10% normal horse serum(NHS) (Gibco) in PBS-T with 0.1% Triton X-100 for 1 h at room temperature to reduce the non-specific binding of the primary antibody.

Techniques: In Vivo, Expressing, In Utero, Electroporation, Construct, Microscopy