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Addgene inc mcherry sept2
Mcherry Sept2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mcherry+sept2/pmCherry-N1_SEPT2+(Plasmid+%2371549)/pm32682163-216-7-8
Average 92 stars, based on 1 article reviews

mcherry sept2 - by Bioz Stars, 2026-09

92/100 stars

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Plasmid Preparation:

Article Title: SEPT7 regulates Ca 2+ entry through Orai channels in human neural progenitor cells and neurons.
Article Snippet: Human neural progenitor cells (hNPCs) are self-renewing cells of neural lineage that can be differentiated into neurons of different subtypes.. Here we show that SEPT7, a member of the family of filament-forming GTPases called septins, prevents constitutive Ca entry through the store-operated Ca entry channel, Orai in hNPCs and in differentiated neurons and is thus required for neuronal calcium homeostasis.. Previous work in Drosophila neurons has shown that loss of one copy of the evolutionarily-conserved dSEPT7 gene leads to elevated Ca entry via Orai, in the absence of ER-Ca store depletion.

Transfection:

Image Search Results

Microtubule-associated SEPT2/6/7 complexes inhibit t he motility of KIF5C, KIF1A, and DDB. A , kymographs show motile ( diagonal lines ) and stationary (vertical lines) KIF5C(1-560)-mCit on an uncoated microtubule ( left ) and a microtubule ( right ) coated with mCherry-SEPT2/6/7 (100 nM). B , mean (±S.D.) landing rates of KIF5C(1-560)-mCit on uncoated microtubules (7.03 ± 1.13 events/μm/min; n = 20 microtubules) and microtubules coated with 100 nM mCherry-SEPT2/6/7 (1.15 ± 0.65 events/μm/min; n = 20 microtubules). ∗∗∗∗ p < 0.0001. C , mean (±S.D.) percentage of KIF5C(1-560)-mCit particles pausing on uncoated microtubules (17.94% ± 16.56%; n = 20 microtubules) and microtubules coated with 100 nM mCherry-SEPT2/6/7 (18.67% ± 16.33%; n = 20 microtubules). n.s., not significant ( p > 0.05). D , mean (±S.D.) velocity of KIF5C(1-560)-mCit particles ( n = 200) on uncoated microtubules (0.78 ± 0.18 μm/s) and microtubules coated with 100 nM of mCherry-SEPT2/6/7 (0.63 ± 0.22 μm/s). ∗∗∗∗ p < 0.0001. E , one-cumulative distribution plot of the run lengths of KIF5C(1-560)-mCit particles ( n = 200) on uncoated microtubules and microtubules with mCherry-SEPT2/6/7 (100 nM). Data were fit to one-phase exponential decay with a decay constant τ (run length), which is shown with the R 2 fit value. The mean (±S.D.) run length values were 1.18 ± 0.52 μm and 0.64 ± 030 μm in the absence and presence of mCherry-SEPT2/6/7, respectively ( p < 0.0001). F , kymographs show motile ( diagonal lines ) and stationary ( vertical lines ) KIF1A(1-393)-GCN4-3XmCit particles on an uncoated microtubule ( left ) and a microtubule ( right ), which was coated with mCherry-SEPT2/6/7 (100 nM). G , mean (±S.D.) landing rates of KIF1A(1-393)-GCN4-3XmCit on uncoated microtubules (5.27 ± 1.62 events/μm/min; n = 20 microtubules) and microtubules coated with 100 nM mCherry-SEPT2/6/7 (1.98 ± 0.79 events/μm/min; n = 20 microtubules). ∗∗∗∗ p < 0.0001. H , mean (±S.D.) percentage of KIF1A(1-393)-GCN4-3XmCit particles pausing on uncoated microtubules (5.99% ± 3.07%; n = 20 microtubules) and microtubules coated with 100 nM mCherry-SEPT2/6/7 (36.19% ± 11.04%; n = 20 microtubules). ∗∗∗∗ p < 0.0001. I , mean (±S.D.) velocity of KIF1A(1-393)-GCN4-3XmCit particles ( n = 200) on uncoated microtubules (1.80 ± 0.34 μm/s) and microtubules coated with mCherry-SEPT2/6/7 (0.80 ± 0.22 μm/s). ∗∗∗∗ p < 0.0001. J , one-cumulative distribution plot of the run lengths of KIF1A(1-393)-GCN4-3XmCit particles ( n = 200) on uncoated microtubules and microtubules with mCherry-SEPT2/6/7 (100 nM). Data were fit to one-phase exponential decay with a decay constant τ (run length), which is shown with the R 2 fit value. The mean (±S.D.) run length values were 6.06 ± 2.85 μm and 2.02 ± 1.13 μm in the absence and presence of mCherry-SEPT2/6/7, respectively ( p < 0.0001). K , kymographs of DDB-GFP motility on an uncoated microtubule and microtubules coated with 20 nM and 50 nM mCherry-SEPT2/6/7. L , mean (±S.D.) landing rates of DDB-GFP particles on uncoated microtubules (0.14 ± 0.09 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) which were coated with 20 nM (0.08 ± 0.06 events/μm/min) and 50 nM mCherry-SEPT2/6/7 (0.02 ± 0.02 events/μm/min). ∗ p = 0.04; ∗∗∗∗ p < 0.0001. M , mean (±S.D.) percentage of DDB-GFP particles that pause on uncoated microtubules (22% ± 27%; n = 20) and microtubules ( n = 20) coated with 20 nM mCherry-SEPT2/6/7 (27% ± 26%). n.s., not significant ( p > 0.05). N , mean (±S.D.) velocity of DDB-GFP particles on uncoated microtubules (0.41 ± 0.27 μm/s; n = 159) and microtubules coated with 20 nM mCherry-SEPT2/6/7 (0.23 ± 0.20 μm/s; n = 150). ∗∗∗∗ p < 0.0001. O , one-cumulative distribution plot of the run lengths of DDB-GFP particles on uncoated microtubules ( n = 159) and microtubules coated with 20 nM mCherry-SEPT2/6/7 ( n = 150). Data were fit to one-phase exponential decay with a decay constant τ (run length) which is shown with the R 2 fit value. The mean (±S.D.) run lengths were 7.38 ± 5.65 μm and 2.57 ± 2.07 μm in the absence and presence of mCherry-SEPT2/6/7, respectively ( p < 0.0001). Statistical analysis of data with normal and non-normal distributions was performed with student's t and Mann-Whitney U tests, respectively. A nonparametric one-way Welch ANOVA test was performed for multiple comparison groups, followed by a post hoc Dunnett T3 test for pairwise comparisons. DDB, dynein-dynactin-bicaudal D.

Journal: The Journal of Biological Chemistry

Article Title: Microtubule-associated septin complexes modulate kinesin and dynein motility with differential specificities

doi: 10.1016/j.jbc.2023.105084

Figure Lengend Snippet: Microtubule-associated SEPT2/6/7 complexes inhibit t he motility of KIF5C, KIF1A, and DDB. A , kymographs show motile ( diagonal lines ) and stationary (vertical lines) KIF5C(1-560)-mCit on an uncoated microtubule ( left ) and a microtubule ( right ) coated with mCherry-SEPT2/6/7 (100 nM). B , mean (±S.D.) landing rates of KIF5C(1-560)-mCit on uncoated microtubules (7.03 ± 1.13 events/μm/min; n = 20 microtubules) and microtubules coated with 100 nM mCherry-SEPT2/6/7 (1.15 ± 0.65 events/μm/min; n = 20 microtubules). ∗∗∗∗ p < 0.0001. C , mean (±S.D.) percentage of KIF5C(1-560)-mCit particles pausing on uncoated microtubules (17.94% ± 16.56%; n = 20 microtubules) and microtubules coated with 100 nM mCherry-SEPT2/6/7 (18.67% ± 16.33%; n = 20 microtubules). n.s., not significant ( p > 0.05). D , mean (±S.D.) velocity of KIF5C(1-560)-mCit particles ( n = 200) on uncoated microtubules (0.78 ± 0.18 μm/s) and microtubules coated with 100 nM of mCherry-SEPT2/6/7 (0.63 ± 0.22 μm/s). ∗∗∗∗ p < 0.0001. E , one-cumulative distribution plot of the run lengths of KIF5C(1-560)-mCit particles ( n = 200) on uncoated microtubules and microtubules with mCherry-SEPT2/6/7 (100 nM). Data were fit to one-phase exponential decay with a decay constant τ (run length), which is shown with the R 2 fit value. The mean (±S.D.) run length values were 1.18 ± 0.52 μm and 0.64 ± 030 μm in the absence and presence of mCherry-SEPT2/6/7, respectively ( p < 0.0001). F , kymographs show motile ( diagonal lines ) and stationary ( vertical lines ) KIF1A(1-393)-GCN4-3XmCit particles on an uncoated microtubule ( left ) and a microtubule ( right ), which was coated with mCherry-SEPT2/6/7 (100 nM). G , mean (±S.D.) landing rates of KIF1A(1-393)-GCN4-3XmCit on uncoated microtubules (5.27 ± 1.62 events/μm/min; n = 20 microtubules) and microtubules coated with 100 nM mCherry-SEPT2/6/7 (1.98 ± 0.79 events/μm/min; n = 20 microtubules). ∗∗∗∗ p < 0.0001. H , mean (±S.D.) percentage of KIF1A(1-393)-GCN4-3XmCit particles pausing on uncoated microtubules (5.99% ± 3.07%; n = 20 microtubules) and microtubules coated with 100 nM mCherry-SEPT2/6/7 (36.19% ± 11.04%; n = 20 microtubules). ∗∗∗∗ p < 0.0001. I , mean (±S.D.) velocity of KIF1A(1-393)-GCN4-3XmCit particles ( n = 200) on uncoated microtubules (1.80 ± 0.34 μm/s) and microtubules coated with mCherry-SEPT2/6/7 (0.80 ± 0.22 μm/s). ∗∗∗∗ p < 0.0001. J , one-cumulative distribution plot of the run lengths of KIF1A(1-393)-GCN4-3XmCit particles ( n = 200) on uncoated microtubules and microtubules with mCherry-SEPT2/6/7 (100 nM). Data were fit to one-phase exponential decay with a decay constant τ (run length), which is shown with the R 2 fit value. The mean (±S.D.) run length values were 6.06 ± 2.85 μm and 2.02 ± 1.13 μm in the absence and presence of mCherry-SEPT2/6/7, respectively ( p < 0.0001). K , kymographs of DDB-GFP motility on an uncoated microtubule and microtubules coated with 20 nM and 50 nM mCherry-SEPT2/6/7. L , mean (±S.D.) landing rates of DDB-GFP particles on uncoated microtubules (0.14 ± 0.09 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) which were coated with 20 nM (0.08 ± 0.06 events/μm/min) and 50 nM mCherry-SEPT2/6/7 (0.02 ± 0.02 events/μm/min). ∗ p = 0.04; ∗∗∗∗ p < 0.0001. M , mean (±S.D.) percentage of DDB-GFP particles that pause on uncoated microtubules (22% ± 27%; n = 20) and microtubules ( n = 20) coated with 20 nM mCherry-SEPT2/6/7 (27% ± 26%). n.s., not significant ( p > 0.05). N , mean (±S.D.) velocity of DDB-GFP particles on uncoated microtubules (0.41 ± 0.27 μm/s; n = 159) and microtubules coated with 20 nM mCherry-SEPT2/6/7 (0.23 ± 0.20 μm/s; n = 150). ∗∗∗∗ p < 0.0001. O , one-cumulative distribution plot of the run lengths of DDB-GFP particles on uncoated microtubules ( n = 159) and microtubules coated with 20 nM mCherry-SEPT2/6/7 ( n = 150). Data were fit to one-phase exponential decay with a decay constant τ (run length) which is shown with the R 2 fit value. The mean (±S.D.) run lengths were 7.38 ± 5.65 μm and 2.57 ± 2.07 μm in the absence and presence of mCherry-SEPT2/6/7, respectively ( p < 0.0001). Statistical analysis of data with normal and non-normal distributions was performed with student's t and Mann-Whitney U tests, respectively. A nonparametric one-way Welch ANOVA test was performed for multiple comparison groups, followed by a post hoc Dunnett T3 test for pairwise comparisons. DDB, dynein-dynactin-bicaudal D.

Article Snippet: The following cotransformations into E. coli BL21 (DE3) (Invitrogen) were performed: His-mCherry-SEPT2 and pnCS SEPT6/7-Strep(+1-57 bp SEPT7 N-term) (SEPT2/6/7), His-mCherry-SEPT5 and SEPT11/7-strep (SEPT5/7/11), and pnEA-vH_His-TEV-SEPT2-mCherry_SEPT6 and pnCS_SEPT7_SEPT9_i1-TEV-Strep (SEPT2/6/7/9).

Techniques: MANN-WHITNEY, Comparison

In microtubule-associated SEPT2/6/7/9 complexes, SEPT9 dampens the i nhibition of KIF1A(1-393) motility by SEPT2/6/7. A , kymographs show motile ( diagonal lines ) and stationary ( vertical lines ) KIF1A(1-393)-GCN4-3XmCit particles on an uncoated microtubule and microtubules which were coated with 10 nM or 100 nM mCherry-SEPT2/6/7/9. B , mean (±S.D.) landing rates of KIF1A(1-393)-GCN4-3XmCit on uncoated microtubules (1.94 ± 0.48 events/μm/min; n = 10 microtubules) and microtubules ( n = 10) coated with 10 nM (1.37 ± 0.35 events/μm/min) or 100 nM mCherry-SEPT2/6/7/9 (0.92 ± 0.37 events/μm/min). ∗∗ p = 0.008; ∗∗∗∗ p < 0.0001. C , mean (±S.D.) percentage of KIF1A(1-393)-GCN4-3XmCit particles that pause on uncoated microtubules (21.09% ± 9.07%; n = 10 microtubules) and microtubules ( n = 10) coated with 10 nM (28.56% ± 12%) or 100 nM mCherry-SEPT2/6/7/9 (48.35% ± 18.82%). n.s., not significant ( p > 0.05); ∗∗∗∗ p < 0.0001. D , mean (±S.D.) velocity of KIF1A(1-393)-GCN4-3XmCit particles ( n = 100) on uncoated microtubules (1.71 ± 0.33 μm/s) and microtubules coated with 10 nM (2.01 ± 0.66 μm/s) or 100 nM mCherry-SEPT2/6/7/9 (1.40 ± 0.37 μm/s). ∗∗ p = 0.002; ∗∗∗∗ p < 0.0001. E , one-cumulative distribution plot of the run lengths of KIF1A(1-393)-GCN4-3XmCit particles ( n = 100) on uncoated microtubules and microtubules coated with 10 nM or 100 nM mCherry-SEPT2/6/7/9. Data were fit to one-phase exponential decay with a decay constant τ (run length) which is shown with the R 2 fit value. The mean (±S.D.) run length value on uncoated microtubules was 7.89 ± 3.88 μm, and the mean (±S.D.) run lengths on microtubules with 10 nM and 100 nM SEPT2/6/7/9 were respectively 5.84 ± 3.18 μm ( p = 0.0003) and 4.44 ± 2.67 μm ( p < 0.0001). Data were statistically analyzed with one-way ANOVA and a post hoc Dunnett test for multiple comparisons ( B and C ) or Kruskal–Wallis test with post hoc Dunn's test for multiple pairwise comparisons ( D and E ). SEPT9, septin 9.

Journal: The Journal of Biological Chemistry

Article Title: Microtubule-associated septin complexes modulate kinesin and dynein motility with differential specificities

doi: 10.1016/j.jbc.2023.105084

Figure Lengend Snippet: In microtubule-associated SEPT2/6/7/9 complexes, SEPT9 dampens the i nhibition of KIF1A(1-393) motility by SEPT2/6/7. A , kymographs show motile ( diagonal lines ) and stationary ( vertical lines ) KIF1A(1-393)-GCN4-3XmCit particles on an uncoated microtubule and microtubules which were coated with 10 nM or 100 nM mCherry-SEPT2/6/7/9. B , mean (±S.D.) landing rates of KIF1A(1-393)-GCN4-3XmCit on uncoated microtubules (1.94 ± 0.48 events/μm/min; n = 10 microtubules) and microtubules ( n = 10) coated with 10 nM (1.37 ± 0.35 events/μm/min) or 100 nM mCherry-SEPT2/6/7/9 (0.92 ± 0.37 events/μm/min). ∗∗ p = 0.008; ∗∗∗∗ p < 0.0001. C , mean (±S.D.) percentage of KIF1A(1-393)-GCN4-3XmCit particles that pause on uncoated microtubules (21.09% ± 9.07%; n = 10 microtubules) and microtubules ( n = 10) coated with 10 nM (28.56% ± 12%) or 100 nM mCherry-SEPT2/6/7/9 (48.35% ± 18.82%). n.s., not significant ( p > 0.05); ∗∗∗∗ p < 0.0001. D , mean (±S.D.) velocity of KIF1A(1-393)-GCN4-3XmCit particles ( n = 100) on uncoated microtubules (1.71 ± 0.33 μm/s) and microtubules coated with 10 nM (2.01 ± 0.66 μm/s) or 100 nM mCherry-SEPT2/6/7/9 (1.40 ± 0.37 μm/s). ∗∗ p = 0.002; ∗∗∗∗ p < 0.0001. E , one-cumulative distribution plot of the run lengths of KIF1A(1-393)-GCN4-3XmCit particles ( n = 100) on uncoated microtubules and microtubules coated with 10 nM or 100 nM mCherry-SEPT2/6/7/9. Data were fit to one-phase exponential decay with a decay constant τ (run length) which is shown with the R 2 fit value. The mean (±S.D.) run length value on uncoated microtubules was 7.89 ± 3.88 μm, and the mean (±S.D.) run lengths on microtubules with 10 nM and 100 nM SEPT2/6/7/9 were respectively 5.84 ± 3.18 μm ( p = 0.0003) and 4.44 ± 2.67 μm ( p < 0.0001). Data were statistically analyzed with one-way ANOVA and a post hoc Dunnett test for multiple comparisons ( B and C ) or Kruskal–Wallis test with post hoc Dunn's test for multiple pairwise comparisons ( D and E ). SEPT9, septin 9.

Techniques:

Microtubule-associated SEPT5/7/11 complexes are permissive to KIF1A motility but inhibit KIF5C and DDB. A , kymographs show motile ( diagonal lines ) and stationary ( vertical lines ) KIF5C(1-560)-mCit on an uncoated microtubule ( left ) and a microtubule ( right ) which was coated with 50 nM mCherry-SEPT5/7/11. Red arrows point to KIF5C(1-560)-mCit motors, which remain immotile with no processive motility prior to dissociation (immotile particles). B , mean (±S.D.) landing rates of KIF5C(1-560)-mCit on uncoated microtubules (2.39 ± 0.53 events/μm/min; n = 20 microtubules) and microtubules coated with 50 nM mCherry-SEPT5/7/11 (1.22 ± 0.33 events/μm/min; n = 20 microtubules). ∗∗∗∗ p < 0.0001. C , mean (±S.D.) percentage of KIF5C(1-560)-mCit particles pausing on uncoated microtubules (6.38% ± 4.00%; n = 20 microtubules) and microtubules coated with 50 nM mCherry-SEPT5/7/11 (11.10% ± 7.50%; n = 20 microtubules). ∗ p = 0.02. D , mean (±S.D.) velocity of KIF5C(1-560)-mCit ( n = 150) on uncoated microtubules (1.19 ± 0.25 μm/s) and microtubules coated with 50 nM of mCherry-SEPT5/7/11 (0.81 ± 0.31 μm/s). ∗∗∗∗ p < 0.0001. E , one-cumulative distribution plot of the run lengths of KIF5C(1-560)-mCit particles ( n = 150) on uncoated microtubules and microtubules with mCherry-SEPT5/7/11 (50 nM). Data were fit to one-phase exponential decay with a decay constant τ (run length), which is shown with the R 2 fit value. The mean (±S.D.) run lengths were 1.58 ± 1 μm and 0.91 ± 0.48 μm in the absence and presence of mCherry-SEPT5/7/11, respectively ( p < 0.0001). F , kymographs show motile ( diagonal lines ) and stationary ( vertical lines ) KIF1A(1-393)-GCN4-3XmCit particles on an uncoated microtubule ( left ) and a microtubule ( right ), which was coated with 50 nM mCherry-SEPT5/7/11. Red arrows point to KIF1A(1-393)-GCN4-3XmCit motors, which associate with a microtubule with no processive motility. G , mean (±S.D.) landing rates of KIF1A(1-393)-GCN4-3XmCit on uncoated microtubules (3.99 ± 1.66 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 50 nM mCherry-SEPT5/7/11 (3.85 ± 1.42 events/μm/min). n.s., not significant ( p > 0.05). H , mean (±S.D.) percentage of KIF1A(1-393)-GCN4-3XmCit particles pausing on uncoated microtubules (13.44% ± 3.69%; n = 20 microtubules) and microtubules ( n = 20) coated with 50 nM mCherry-SEPT5/7/11 (19.33% ± 5.63%; n = 20). ∗∗∗ p = 0.0004. I , mean (±S.D.) velocity of KIF1A(1-393)-GCN4-3XmCit particles ( n = 148) on uncoated microtubules (1.69 ± 0.38 μm/s) and microtubules coated with 50 nM mCherry-SEPT5/7/11 (1.32 ± 0.33 μm/s). ∗∗∗∗ p < 0.0001. J , one-cumulative distribution plot of the run lengths of KIF1A(1-393)-GCN4-3XmCit particles ( n = 148) on uncoated microtubules and microtubules with mCherry-SEPT5/7/11 (50 nM). Data were fit to one-phase exponential decay with a decay constant τ (run length), which is shown with the R 2 fit value. The mean (±S.D.) run length values were 5.25 ± 2.59 μm and 4.68 ± 2.63 μm in the absence and presence of mCherry-SEPT2/6/7, respectively ( p > 0.05). K , kymographs of DDB-GFP on an uncoated microtubule and microtubules which were coated with 10 nM and 50 nM mCherry-SEPT5/7/11. Red arrows point to DDB-GFP particles, which associate with microtubules with no processive motility (immotile particles). L , mean (±S.D.) landing rates of DDB-GFP particles on uncoated microtubules (0.26 ± 0.15 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 10 nM (0.19 ± 0.14 events/μm/min) and 50 nM mCherry-SEPT5/7/11 (0.01 ± 0.02 events/μm/min). n.s., not significant ( p > 0.05); ∗∗∗∗ p < 0.0001. M , mean (±S.D.) percentage of DDB-GFP particles that pause on uncoated microtubules (11.85% ± 14.98%; n = 20 microtubules) and microtubules ( n = 20) which were coated with 10 nM mCherry-SEPT5/7/11 (28.37% ± 24.35%) ∗ p = 0.01. N , mean (±S.D.) velocity of DDB-GFP particles on uncoated microtubules (0.42 ± 0.28 μm/s; n = 150) and microtubules coated with 50 nM mCherry-SEPT5/7/11 (0.28 ± 0.22 μm/s; n = 155). ∗∗∗∗ p < 0.0001. O , one-cumulative distribution plot of the run lengths of DDB-GFP ( n = 150–155) on uncoated microtubules and microtubules coated with mCherry-SEPT5/7/11 (50 nM). Data were fit to one-phase exponential decay with a decay constant τ (run length), which is shown with the R 2 fit value. The mean (±S.D.) run length values were 6.29 ± 4.86 μm ( n = 150) and 3.14 ± 2.80 μm in the absence and presence of mCherry-SEPT5/7/11, respectively ( p < 0.0001). Statistical analysis of data with normal and non-normal distributions was performed with student's t and Mann-Whitney U tests, respectively. A nonparametric one-way ANOVA Kruskal–Wallis test was performed for multiple comparison groups, followed by a post hoc Dunn's test for pairwise comparisons. DDB, dynein-dynactin-bicaudal D.

Journal: The Journal of Biological Chemistry

Article Title: Microtubule-associated septin complexes modulate kinesin and dynein motility with differential specificities

doi: 10.1016/j.jbc.2023.105084

Figure Lengend Snippet: Microtubule-associated SEPT5/7/11 complexes are permissive to KIF1A motility but inhibit KIF5C and DDB. A , kymographs show motile ( diagonal lines ) and stationary ( vertical lines ) KIF5C(1-560)-mCit on an uncoated microtubule ( left ) and a microtubule ( right ) which was coated with 50 nM mCherry-SEPT5/7/11. Red arrows point to KIF5C(1-560)-mCit motors, which remain immotile with no processive motility prior to dissociation (immotile particles). B , mean (±S.D.) landing rates of KIF5C(1-560)-mCit on uncoated microtubules (2.39 ± 0.53 events/μm/min; n = 20 microtubules) and microtubules coated with 50 nM mCherry-SEPT5/7/11 (1.22 ± 0.33 events/μm/min; n = 20 microtubules). ∗∗∗∗ p < 0.0001. C , mean (±S.D.) percentage of KIF5C(1-560)-mCit particles pausing on uncoated microtubules (6.38% ± 4.00%; n = 20 microtubules) and microtubules coated with 50 nM mCherry-SEPT5/7/11 (11.10% ± 7.50%; n = 20 microtubules). ∗ p = 0.02. D , mean (±S.D.) velocity of KIF5C(1-560)-mCit ( n = 150) on uncoated microtubules (1.19 ± 0.25 μm/s) and microtubules coated with 50 nM of mCherry-SEPT5/7/11 (0.81 ± 0.31 μm/s). ∗∗∗∗ p < 0.0001. E , one-cumulative distribution plot of the run lengths of KIF5C(1-560)-mCit particles ( n = 150) on uncoated microtubules and microtubules with mCherry-SEPT5/7/11 (50 nM). Data were fit to one-phase exponential decay with a decay constant τ (run length), which is shown with the R 2 fit value. The mean (±S.D.) run lengths were 1.58 ± 1 μm and 0.91 ± 0.48 μm in the absence and presence of mCherry-SEPT5/7/11, respectively ( p < 0.0001). F , kymographs show motile ( diagonal lines ) and stationary ( vertical lines ) KIF1A(1-393)-GCN4-3XmCit particles on an uncoated microtubule ( left ) and a microtubule ( right ), which was coated with 50 nM mCherry-SEPT5/7/11. Red arrows point to KIF1A(1-393)-GCN4-3XmCit motors, which associate with a microtubule with no processive motility. G , mean (±S.D.) landing rates of KIF1A(1-393)-GCN4-3XmCit on uncoated microtubules (3.99 ± 1.66 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 50 nM mCherry-SEPT5/7/11 (3.85 ± 1.42 events/μm/min). n.s., not significant ( p > 0.05). H , mean (±S.D.) percentage of KIF1A(1-393)-GCN4-3XmCit particles pausing on uncoated microtubules (13.44% ± 3.69%; n = 20 microtubules) and microtubules ( n = 20) coated with 50 nM mCherry-SEPT5/7/11 (19.33% ± 5.63%; n = 20). ∗∗∗ p = 0.0004. I , mean (±S.D.) velocity of KIF1A(1-393)-GCN4-3XmCit particles ( n = 148) on uncoated microtubules (1.69 ± 0.38 μm/s) and microtubules coated with 50 nM mCherry-SEPT5/7/11 (1.32 ± 0.33 μm/s). ∗∗∗∗ p < 0.0001. J , one-cumulative distribution plot of the run lengths of KIF1A(1-393)-GCN4-3XmCit particles ( n = 148) on uncoated microtubules and microtubules with mCherry-SEPT5/7/11 (50 nM). Data were fit to one-phase exponential decay with a decay constant τ (run length), which is shown with the R 2 fit value. The mean (±S.D.) run length values were 5.25 ± 2.59 μm and 4.68 ± 2.63 μm in the absence and presence of mCherry-SEPT2/6/7, respectively ( p > 0.05). K , kymographs of DDB-GFP on an uncoated microtubule and microtubules which were coated with 10 nM and 50 nM mCherry-SEPT5/7/11. Red arrows point to DDB-GFP particles, which associate with microtubules with no processive motility (immotile particles). L , mean (±S.D.) landing rates of DDB-GFP particles on uncoated microtubules (0.26 ± 0.15 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 10 nM (0.19 ± 0.14 events/μm/min) and 50 nM mCherry-SEPT5/7/11 (0.01 ± 0.02 events/μm/min). n.s., not significant ( p > 0.05); ∗∗∗∗ p < 0.0001. M , mean (±S.D.) percentage of DDB-GFP particles that pause on uncoated microtubules (11.85% ± 14.98%; n = 20 microtubules) and microtubules ( n = 20) which were coated with 10 nM mCherry-SEPT5/7/11 (28.37% ± 24.35%) ∗ p = 0.01. N , mean (±S.D.) velocity of DDB-GFP particles on uncoated microtubules (0.42 ± 0.28 μm/s; n = 150) and microtubules coated with 50 nM mCherry-SEPT5/7/11 (0.28 ± 0.22 μm/s; n = 155). ∗∗∗∗ p < 0.0001. O , one-cumulative distribution plot of the run lengths of DDB-GFP ( n = 150–155) on uncoated microtubules and microtubules coated with mCherry-SEPT5/7/11 (50 nM). Data were fit to one-phase exponential decay with a decay constant τ (run length), which is shown with the R 2 fit value. The mean (±S.D.) run length values were 6.29 ± 4.86 μm ( n = 150) and 3.14 ± 2.80 μm in the absence and presence of mCherry-SEPT5/7/11, respectively ( p < 0.0001). Statistical analysis of data with normal and non-normal distributions was performed with student's t and Mann-Whitney U tests, respectively. A nonparametric one-way ANOVA Kruskal–Wallis test was performed for multiple comparison groups, followed by a post hoc Dunn's test for pairwise comparisons. DDB, dynein-dynactin-bicaudal D.

Techniques: MANN-WHITNEY, Comparison

SEPT5/7/11 promotes DDB and kinesin tethering to microtubules. A , mean (±S.D.) number of immotile DDB-GFP events per micrometer of uncoated microtubules (0.07 ± 0.06 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 10 nM (0.15 ± 0.07 events/μm/min) or 50 nM mCherry-SEPT5/7/11 (0.28 ± 0.12 events/μm/min). ∗∗ p = 0.006; ∗∗∗∗ p < 0.0001. B , mean (±S.D.) number of immotile DDB-GFP events per micrometer of uncoated microtubules (0.05 ± 0.05 events/μm/min; n = 20 microtubules) and microtubules (n = 20) coated with 20 nM (0.07 ± 0.05 events/μm/min) or 50 nM mCherry-SEPT2/6/7 (0.06 ± 0.04 events/μm/min). n.s., not significant ( p > 0.05). C , mean (±S.D.) number of immotile KIF5C(1-560)-mCit particles per micrometer of uncoated microtubules (0.18 ± 0.12 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 50 nM mCherry-SEPT5/7/11 (0.37 ± 0.15 events/μm/min). ∗∗∗∗ p < 0.0001. D , mean (±S.D.) number of immotile KIF5C(1-560)-mCit events per micrometer of uncoated microtubules (1.12 ± 0.41 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 100 nM mCherry-SEPT2/6/7 (1.23 ± 0.49 events/μm/min). n.s., not significant ( p > 0.05). E , mean (±SEM) number of immotile KIF1A(1-393)-GCN4-3XmCit events per micrometer of uncoated microtubule (0.42 ± 0.24 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 50 nM mCherry-SEPT5/7/11 (0.57 ± 0.18 events/μm). ∗ p = 0.03. F , mean (±SEM) number of immotile KIF1A(1-393)-GCN4-3XmCit events per micrometer of uncoated microtubule (0.75 ± 0.39 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 50 nM mCherry-SEPT2/6/7 (0.67 ± 0.29 events/μm/min). n.s., not significant ( p > 0.05). Statistical analysis of data with normal and non-normal distributions was performed with student's t and Mann-Whitney U tests, respectively. A nonparametric one-way ANOVA Kruskal–Wallis test was performed for multiple comparison groups, followed by a post hoc Dunn's test for pairwise comparisons. DDB, dynein-dynactin-bicaudal D.

Journal: The Journal of Biological Chemistry

Article Title: Microtubule-associated septin complexes modulate kinesin and dynein motility with differential specificities

doi: 10.1016/j.jbc.2023.105084

Figure Lengend Snippet: SEPT5/7/11 promotes DDB and kinesin tethering to microtubules. A , mean (±S.D.) number of immotile DDB-GFP events per micrometer of uncoated microtubules (0.07 ± 0.06 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 10 nM (0.15 ± 0.07 events/μm/min) or 50 nM mCherry-SEPT5/7/11 (0.28 ± 0.12 events/μm/min). ∗∗ p = 0.006; ∗∗∗∗ p < 0.0001. B , mean (±S.D.) number of immotile DDB-GFP events per micrometer of uncoated microtubules (0.05 ± 0.05 events/μm/min; n = 20 microtubules) and microtubules (n = 20) coated with 20 nM (0.07 ± 0.05 events/μm/min) or 50 nM mCherry-SEPT2/6/7 (0.06 ± 0.04 events/μm/min). n.s., not significant ( p > 0.05). C , mean (±S.D.) number of immotile KIF5C(1-560)-mCit particles per micrometer of uncoated microtubules (0.18 ± 0.12 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 50 nM mCherry-SEPT5/7/11 (0.37 ± 0.15 events/μm/min). ∗∗∗∗ p < 0.0001. D , mean (±S.D.) number of immotile KIF5C(1-560)-mCit events per micrometer of uncoated microtubules (1.12 ± 0.41 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 100 nM mCherry-SEPT2/6/7 (1.23 ± 0.49 events/μm/min). n.s., not significant ( p > 0.05). E , mean (±SEM) number of immotile KIF1A(1-393)-GCN4-3XmCit events per micrometer of uncoated microtubule (0.42 ± 0.24 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 50 nM mCherry-SEPT5/7/11 (0.57 ± 0.18 events/μm). ∗ p = 0.03. F , mean (±SEM) number of immotile KIF1A(1-393)-GCN4-3XmCit events per micrometer of uncoated microtubule (0.75 ± 0.39 events/μm/min; n = 20 microtubules) and microtubules ( n = 20) coated with 50 nM mCherry-SEPT2/6/7 (0.67 ± 0.29 events/μm/min). n.s., not significant ( p > 0.05). Statistical analysis of data with normal and non-normal distributions was performed with student's t and Mann-Whitney U tests, respectively. A nonparametric one-way ANOVA Kruskal–Wallis test was performed for multiple comparison groups, followed by a post hoc Dunn's test for pairwise comparisons. DDB, dynein-dynactin-bicaudal D.

Techniques: MANN-WHITNEY, Comparison

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