Control of endothelial cell polarity and sprouting angiogenesis by non-centrosomal microtubules
- Utrecht University, Netherlands
- University of Liège, Belgium
Figures
Figure 1 with 2 supplements
The centrosome is not essential for angiogenic migration and sprouting.
(A) Imaging of control or centrinone-treated HUVECs stained for MT (α-tubulin, cyan hot) using STED microscopy. Arrow points toward the centrosome and the plot shows the average fluorescence …
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Figure 1—source data 1
An Excel sheet with numerical data on the quantification of the effect of centrinone treatment on the EC mean intensity of α-tubulin signal, MT dynamics parameters, EB comet number, the polarization of Golgi during migration, the efficiency of wound closure, the cumulative length of spheroid sprouts and the proportion of sprouting ECs with centrosome as well as the effect of CPAP depletion on the cumulative length of spheroid sprouts represented as plots in Figure 1A–F,H.
- https://doi.org/10.7554/eLife.33864.006
Figure 1—figure supplement 1
The centrosome is not essential for angiogenic migration and sprouting.
(A) Staining of HUVECs during wound healing assay (2D, wide-field fluorescence image) and spheroid sprouting (3D, Z-maximum projection of confocal fluorescence images) for the centriolar marker …
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Figure 1—figure supplement 1—source data 1
An Excel sheet with numerical data on the quantification of the polarization of Golgi and centrosome during 2D migration and 3D sprouting as well as of centrosome removal efficiency using different (peri)centriolar markers, Golgi area and dispersion and MT nucleation activity and EB3 Golgi enrichment after nocodazole washout after centrinone treatment represented as plots (or as mean value ± SD for 1B) in Figure 1—figure supplement 1A–D.
- https://doi.org/10.7554/eLife.33864.007
Figure 1—figure supplement 2
The centrosome is not essential for angiogenic migration and sprouting.
(A) Western blots of total extract from HUVECs treated or not treated with centrinone were used to quantify the levels of α-tubulin and CAMSAP2 protein expression as well as of tubulin …
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Figure 1—figure supplement 2—source data 1
An Excel sheet with numerical data on the quantification of the effect of centrinone treatment on the expression of CAMSAP2 and various post-translationally modified tubulin in ECs, the mean intensity of acetylated tubulin signal, the density of CAMSAP2 stretches, the intensity of VE-Cadherin and ZO-1 signal at cell junctions, the velocity and directionality of cell migration during scratch-wound assays, as well as the effect of CPAP depletion on centrosome removal efficiency and the proportion of 3D sprouting ECs with centrosome represented as plots in Figure 1—figure supplement 2A–G.
- https://doi.org/10.7554/eLife.33864.008
Figure 2 with 2 supplements
CAMSAP2 is required for maintaining non-centrosomal MTs and EC migration.
(A) Staining of CAMSAP2 (white) and α-tubulin (red) in serum-starved HUVECs before or after a 2 hr treatment with VEGF. Wide-field fluorescence images are shown. (B) Western blots of total extracts …
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Figure 2—source data 1
An Excel sheet with numerical data on the quantification of the effect of CAMSAP2 silencing on MT dynamics parameters, the efficiency of wound closure, the cumulative length of spheroid sprouts, their number and average length, and the cumulative length of spheroid sprouts re-expressing CAMSAP2 represented as plots in Figure 2D,E,G,H.
- https://doi.org/10.7554/eLife.33864.012
Figure 2—figure supplement 1
CAMSAP2 is required for maintaining non-centrosomal MTs and cell migration in ECs.
(A) Plots showing the number per cell and length of CAMSAP2 stretches in HUVECs treated and stained as in Figure 2A, n = 30 cells per condition. (B) Western blots of total extracts of HUVECs 48 hr …
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Figure 2—figure supplement 1—source data 1
An Excel sheet with numerical data on the quantification of CAMSAP2 stretch number and length after VEGF treatment, as well as the effect of CAMSAP2 depletion on the EC mean intensity of α-tubulin signal, EB comet number, the expression of CAMSAP2 and various post-translationally modified tubulin, MT nucleation activity and EB3 Golgi and centrosome enrichment after nocodazole washout, the number, speed and length of KIF13B tracks and the velocity of cell migration during scratch-wound assays represented as plots in Figure 2—figure supplement 1A,C,E,F,H,I.
- https://doi.org/10.7554/eLife.33864.013
Figure 2—figure supplement 2
CAMSAP2 is required for maintaining non-centrosomal MTs and cell migration in ECs.
(A) Quantification of mitotic index, based on a phalloidin/DAPI staining, and of doubling time, based on manual counting with trypan blue, 48 hr after transfection with the indicated siRNAs in three …
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Figure 2—figure supplement 2—source data 1
An Excel sheet with numerical data on the quantification of the EC mitotic index and doubling time after CAMSAP2 depletion, the cumulative length of spheroid sprouts after CAMSAP2 depletion and treatment with thymidine and after CAMSAP2 and CAMSAP3 depletion represented as plot in Figure 2—figure supplement 2A,B,D.
- https://doi.org/10.7554/eLife.33864.014
Figure 3 with 1 supplement
CAMSAP2 is required for stabilization of one major cell protrusion.
(A) Staining for F-actin (phalloidin, green), α-tubulin (red) and DNA (DAPI, blue) in control or CAMSAP2-depleted sprouting spheroids. Z-maximum projections of confocal images are shown. (B,C) …
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Figure 3—source data 1
An Excel sheet with numerical data on the quantification of the effect of CAMSAP2 depletion on the 3D elongation of ECs, the number of their 3D protrusions and the length of the longest one, their polarity index (protrusion organization), the persistence of the protrusions over time and the enrichment of α-tubulin signal in the longest protrusion represented as plots in Figure 3B,C,D,F,G.
- https://doi.org/10.7554/eLife.33864.017
Figure 3—figure supplement 1
CAMSAP2 is required for stabilization of one major cell protrusion.
(A) Bright-field micrographs of a time-course sprouting experiment from spheroids of control or CAMSAP2-depleted HUVECs and quantification of the number and length of the sprouts, n = 20 spheroids …
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Figure 3—figure supplement 1—source data 1
An Excel sheet with numerical data on the quantification of the effect of CAMSAP2 depletion on the number and length of spheroid protrusions over time, the total and average length of the 3D protrusions of isolated ECs, the binning of the average protrusion length by their direction, the polarity index of the 3D protrusions in relation to their length and the number of 3D protrusions over time represented as plots in Figure 3—figure supplement 1A,C,E–G.
- https://doi.org/10.7554/eLife.33864.018
Figure 4 with 2 supplements
CAMSAP2 depletion phenotypes cannot be explained by changes in the actin cytoskeleton and cell contractility.
(A) Imaging of control or CAMSAP2 siRNA-treated HUVECs during 2D wound healing assay stained for F-actin (phalloidin, cyan hot) using STED microscopy. The plot shows the average fluorescence …
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Figure 4—source data 1
An Excel sheet with numerical data on the quantification of the effect of CAMSAP2 depletion on the intensity of phalloidin signal in 2D (mean intensity) and in 3D (maximum intensity) ECs, as well as on the EC polarity index, the persistence of the protrusions over time and the cumulative length of spheroid sprouts after Y27632 and blebbistatin treatment represented as plots in Figure 4A,C–F.
- https://doi.org/10.7554/eLife.33864.022
Figure 4—figure supplement 1
CAMSAP2 depletion phenotypes cannot be explained by changes in the actin cytoskeleton and cell contractility.
(A) Phalloidin stainings of control or CAMSAP2 siRNA-treated HUVECs shown in Figure 4A were used to quantify the width of the lamellipodia, the percentage of the leading edge covered with …
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Figure 4—figure supplement 1—source data 1
An Excel sheet with numerical data on the quantification of the effect of CAMSAP2 depletion on the proportion of coverage and the width of lamellipodia, the cumulative length and the width of stress fibers in 2D migrationg ECs, the activation level of Rho and Rac1 GTPases, the intensity of VE-Cadherin and ZO-1 signal at cell junctions and the intensity of phalloidin signal in 3D represented as plots (or mean value ± SD for 1B) in Figure 4—figure supplement 1A–D.
- https://doi.org/10.7554/eLife.33864.023
Figure 4—figure supplement 2
CAMSAP2 depletion phenotypes cannot be explained by changes in actin cytoskeleton and cell contractility.
(A–E) Staining for F-actin in 3D cultured control or CAMSAP2-inactivated HUVECs treated or not treated with Y-27632 or blebbistatin (as in Figure 4D) were used to create binary cell masks using …
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Figure 4—figure supplement 2—source data 1
An Excel sheet with numerical data on the quantification of the effect of CAMSAP2 depletion and Y632 or blebbistatin treatment on the cumulative and average length of 3D protrusions from isolated EC, their number and the length of the longest ones as well as on the number and length of spheroid sprouts represented as plot in Figure 4—figure supplement 2B–F.
- https://doi.org/10.7554/eLife.33864.024
Figure 5 with 1 supplement
CAMSAP2 participates in Golgi polarization during 2D migration and 3D sprouting.
(A) Directionality of cell movement (distance between the start and end point of migration divided by the total distance travelled) during a phase-contrast time-lapse recording of a wound healing …
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Figure 5—source data 1
An Excel sheet with numerical data on the quantification of the effect of CAMSAP2 depletion on the directionality of EC migration, the correlation between the position of the lamellipodia, Golgi and wound during migration, the proportion of Rab6 tracks in the front of migrating ECs, the polarization of Golgi in sprouting ECs and the proportion of Rab6 tracks (anterograde and retrograde) in the 3D longest protrusion represented as plots in Figure 5A–E.
- https://doi.org/10.7554/eLife.33864.027
Figure 5—figure supplement 1
CAMSAP2 participates in Golgi polarization during 2D migration and 3D sprouting.
(A) Staining for γ-tubulin (green), Golgi (GM130, red) and DNA (DAPI, blue) in HUVECs transfected with control or CAMSAP2 siRNAs during a 2D wound healing assay. Z-maximum projections of confocal …
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Figure 5—figure supplement 1—source data 1
An Excel sheet with numerical data on the quantification of the effect of CAMSAP2 depletion on the polarization of the centrosome during migration and the speed, duration and length of Rab6 tracks in 2D migrating ECs represented as plots in Figure 5—figure supplement 1A,B.
- https://doi.org/10.7554/eLife.33864.028
Figure 6 with 1 supplement
Loss of non-centrosomal MTs has a more severe impact than their detachment from the Golgi in 3D but not in 2D.
(A) Staining for CAMSAP2 (white, green) and Golgi (GM130, red) in HUVECs transfected with the indicated siRNA. The plot shows CAMSAP2 enrichment at the Golgi (ratio between the average CAMSAP2 …
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Figure 6—source data 1
An Excel sheet with numerical data on the quantification of the enrichment of CAMSAP2 at the Golgi, the directionality and efficiency of migration during scratch-wound assays and the cumulative length of spheroid sprouts in the absence of AKAP450, MMG or CAMSAP2 represented as plots in Figure 6A–C.
- https://doi.org/10.7554/eLife.33864.031
Figure 6—figure supplement 1
Loss of non-centrosomal MTs has a more severe impact than their detachment from the Golgi in 3D but not in 2D.
(A) Western blots of HUVEC extracts 72 hr after transfection with control, AKAP450 or MMG siRNAs using antibodies against AKAP450 or MMG, and Ku80 as loading control. (B) Normalized fluorescence …
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Figure 6—figure supplement 1—source data 1
An Excel sheet with numerical data on the quantification of the profile of CAMSAP2 and GM130 signal along the cell radius, the velocity of migration and the polarization of Golgi during scratch-wound assays in the absence of AKAP450, MMG or CAMSAP2 represented as plots in Figure 6—figure supplement 1B–D.
- https://doi.org/10.7554/eLife.33864.032
Figure 7 with 1 supplement
The centrosome inhibits cell polarization and sprouting in the absence of CAMSAP2.
(A,B) Staining for Golgi (GM130, red) and α-tubulin (white) in control and centrinone-treated HUVECs transfected with the indicated siRNAs. Z-maximum projections of confocal images (A) and average …
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Figure 7—source data 1
An Excel sheet with numerical data on the quantification of the mean intensity of EC α-tubulin signal, the enrichment of γ- and α-tubulin at the Golgi and the cumulative length of spheroid sprouts in the absence of AKAP450, MMG or CAMSAP2 and after centrinone treatment represented as plots in Figure 7B,D,E.
- https://doi.org/10.7554/eLife.33864.035
Figure 7—figure supplement 1
The centrosome inhibits cell polarization and sprouting in the absence of CAMSAP2.
(A) EB3 staining in control and centrinone-treated HUVECs transfected with the indicated siRNAs was used to quantify the density of EB comets, n = 19, 20, 20, 21, 20, 20, 20, 21 cells, histogram …
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Figure 7—figure supplement 1—source data 1
An Excel sheet with numerical data on the quantification of the number of EB comets in 2D ECs and the enrichment of EB at the Golgi after nocodazole washout in the absence of AKAP450, MMG or CAMSAP2 together with centrinone treatment represented as plots in Figure 7—figure supplement 1A,B.
- https://doi.org/10.7554/eLife.33864.036
Figure 8 with 1 supplement
Non-centrosomal MTs are required to create protrusion asymmetry.
(A) Imaging of control and centrinone-treated HUVECs transfected with the indicated siRNAs and stained for MTs (α-tubulin, cyan hot) using STED microscopy. MT images were split into a radial and …
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Figure 8—source data 1
An Excel sheet with numerical data on the quantification of the effect of MMG or CAMSAP2 depletion together with centrinone treatment on the proportion of the non-radial MT network, the EC polarity index and the enrichment of α-tubulin signal in the 3D longest protrusion as well as the effect of MMG depletion and centrinone treatment on the enrichment of CAMSAP2 at the Golgi and in the longest protrusion in 3D represented as plots in Figure 8A–D.
- https://doi.org/10.7554/eLife.33864.039
Figure 8—figure supplement 1
Non-centrosomal MTs are required to create protrusion asymmetry.
Quantification of 3D morphology of HUVECs treated as indicated, using ImageJ; plots show the total number of protrusions per cell and the length of the longest protrusion, n = 44, 36, 35, 38, 36 and …
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Figure 8—figure supplement 1—source data 1
An Excel sheet with numerical data on the quantification of the effect of MMG or CAMSAP2 depletion together with centrinone treatment on the number and length of the 3D protrusions of isolated ECs represented as plot in Figure 8—figure supplement 1.
- https://doi.org/10.7554/eLife.33864.040
Figure 9 with 1 supplement
CAMSAP2 plays a role in sprouting angiogenesis in vivo.
(A) Live confocal images (Z-maximum projections) of 48 hpf Tg(Fli1ep:Lifeact-EGFP) embryos injected with control or CAMSAP2b morpholinos. Arterial and venous intersegmental vessels are indicated by …
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Figure 9—source data 1
An Excel sheet with numerical data on the quantification of the effect of CAMSAP2b inactivation and its re-expression (for 1B) in zebrafish on the proportion of venous intersegmental vessels, the length of venous sprouts over time, the variation of their length and direction over time and the growth and directional persistence of secondary sprout formation represented as plots in Figure 9B,D–F.
- https://doi.org/10.7554/eLife.33864.043
Figure 9—figure supplement 1
CAMSAP2 plays a role in sprouting angiogenesis in vivo.
(A) RT-PCR analysis of Tg(fli1a:eGFP) embryos injected with a splice-blocking morpholino targeting the exon2/intron2 boundary in Camsap2b (black box), a control or no morpholino (-) with primers …
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Figure 9—figure supplement 1—source data 1
An Excel sheet with numerical data on the quantification of the efficiency of CAMSAP2b silencing, the number of secondary venous sprouts at 34 and 48 hpf and the number of loops in the caudal vein plexus after CAMSAP2b-directed morpholino injection in zebrafish embryos as well as of the number of secondary venous sprouts at 36 hpf after re-expression of CAMSAP2 represented as plots in Figure 9—figure supplement 1B,D,F,G.
- https://doi.org/10.7554/eLife.33864.044
Videos
Video 1
Time-lapse imaging of directional venous sprouting in control Tg(Fli1ep:Lifeact-EGFP) embryos.
Time is hr: min post-fertilization. Z-series images in the region centered on the yolk extension end using a 2-µm-step confocal based scan covering 70 µm depth were taken every 10 min. This video …
Video 2
Time-lapse imaging of venous sprouting, arterial intersegmental vessel fusion and parachordal lymphangioblast assembly in control Tg(Fli1ep:Lifeact-EGFP) embryos.
Time is hr: min post-fertilization. This video was acquired as described in Video 1 and illustrates the two different outcomes of venous sprouting: arterial intersegmental vessel fusion or …
Video 3
Time-lapse imaging of unstable venous sprouting in CAMSAP2 morphant Tg(Fli1ep:Lifeact-EGFP) embryos.
Time is hr: min post-fertilization. This video was acquired as described in Video 1 and shows two highly unstable venous sprouts.
Video 4
Time-lapse imaging of non-persistent venous sprouting in CAMSAP2 morphant Tg(Fli1ep:Lifeact-EGFP) embryos.
Time is hr: min post-fertilization. This video was acquired as described in Video 1 and illustrates the instability and lack of directional persistence of venous sprouting.
Video 5
Time-lapse imaging of venous sprouting in CAMSAP2 morphant Tg(Fli1ep:Lifeact-EGFP) embryos.
Time is hr: min post-fertilization. This video was acquired as described in Video 1. The rightmost arterial intersegmental vessel exhibits atypical fusion with two distinct venous sprouts.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Dario rerio) | Tg(fli1a:eGFP)y1 | Zebrafish facility GIGA institute, Liege University | ID_Zfin:ZDB-TGCONSTRCT-070117–94 | |
| Strain, strain background (D. rerio) | Tg(Fli1ep:Lifeact-EGFP) | Zebrafish facility GIGA institute, Liege University;Phng et al. (2013); PMID: 24046319 | ID_Zfin:ZDB-TGCONSTRCT-140610–8 | |
| Cell line (Homo sapiens) | HUVECs | Lonza | Lonza:C2519AS | Primary endothelial cells cultured as recommended by Lonza |
| Antibody | anti-CAMSAP2 (rabbit polyclonal) | Novus | Novus:NBP1-21402; RRID:AB_1659977 | (1:200) for IF; (1:1000) for WB |
| Antibody | anti-CEP135 (rabbit polyclonal) | Sigma-Aldrich | Sigma-Aldrich:SAB4503685; RRID:AB_10746232 | (1:300) |
| Antibody | anti-acetylated tubulin (rabbit polyclonal) | Sigma-Aldrich | Sigma-Aldrich:T7451; RRID:AB_609894 | (1:300) |
| Antibody | anti-polyglutamylated tubulin (rabbit polyclonal) | Sigma-Aldrich | Sigma-Aldrich:T9822; RRID:AB_477598 | (1:2000) |
| Antibody | anti- detyrosinated tubulin (rabbit polyclonal) | Abcam | Abcam:ab48389; RRID:AB_869990 | (1:2000) |
| Antibody | anti-γtubulin (rabbit polyclonal) | Sigma-Aldrich | Sigma-Aldrich:T3559, RRID:AB_477575 | (1:300) for IF; (1:1000) for WB |
| Antibody | anti-CDK5RAP2 (rabbit polyclonal) | Bethyl Laboratories | Bethyl Laboratories:A300-554A | (1:500) |
| Antibody | anti-EB3 (rabbit polyclonal) | Stepanova et al., 2003; PMID: 12684451 | (1:400) | |
| Antibody | anti-MMG8 (rabbit polyclonal) | Wang et al. (2014); PMID: 25217626 | (1:300) for IF; (1:1000) for WB | |
| Antibody | anti-MYOSIN IIb (goat polyclonal) | Santa-Cruz biotechnology | Santa-Cruz biotechnology:sc-47205; RRID:AB_2297998 | (1:200) |
| Antibody | anti-PCM1 (goat polyclonal) | Santa-Cruz biotechnology | Santa-Cruz biotechnology:sc-50164; RRID:AB_2160195 | (1:300) |
| Antibody | anti-GM130 (mouse monoclonal) | BD Biosciences | BD Biosciences:610823; RRID:AB_398142 | (1:600) |
| Antibody | anti-pericentrin (mouse monoclonal) | BD Biosciences | BD Biosciences:611815; RRID:AB_399295 | (1:300) |
| Antibody | anti-EB1 (mouse monoclonal) | BD Biosciences | BD Biosciences:610535; RRID:AB_397892 | (1:400) |
| Antibody | anti-VE-Cadherin (mouse monoclonal) | BD Biosciences | BD Biosciences:610252; RRID:AB_2276073 | (1:500) |
| Antibody | anti-ZO-1 (mouse monoclonal) | BD Biosciences | BD Biosciences:610966; RRID:AB_398279 | (1:200) |
| Antibody | anti-AKAP450 (mouse monoclonal) | BD Biosciences | BD Biosciences:611518; RRID:AB_398978 | (1:300) for IF; (1:500) for WB |
| Antibody | anti-KU80 (mouse monoclonal) | BD Biosciences | BD Biosciences:611360; RRID:AB_398882 | (1:3000) |
| Antibody | anti-CAMSAP3 (mouse monoclonal) | Sigma-Aldrich | Sigma-Aldrich:SAB4200415 | (1:500) |
| Antibody | anti-αtubulin (mouse monoclonal) | Sigma-Aldrich | Sigma-Aldrich:T5168; RRID:AB_477579 | (1:400) for IF; (1:2000) for WB |
| Antibody | anti-γtubulin (mouse monoclonal) | Sigma-Aldrich | Sigma-Aldrich: T6557; RRID:AB_477584 | (1:300) |
| Antibody | anti-NEDD1 (mouse monoclonal) | Abnova | Abnova:H00121441-M05; RRID:AB_534956 | (1:300) |
| Antibody | anti-αtubulin YL1/2 (rat monoclonal) | Pierce | Pierce: MA1-80017; RRID:AB_2210201 | (1:400) |
| Antibody | anti-CPAP (rabbit polyclonal) | Kohlmaier et al. (2009); PMID: 19481460 | (1:200) | |
| Antibody | Alexa Fluor 488-, 594- and 647- secondaries | Molecular Probes | (1:400) | |
| Antibody | Alexa Fluor 488-,and 594- phalloidin | Molecular Probes | (1:500) | |
| Antibody | Abberior STAR 635P- anti-mouse | Sigma-Aldrich | Sigma-Aldrich:2-0002-007-5 | (1:200) |
| Antibody | Atto 647N Phalloidin | Sigma-Aldrich | Sigma-Aldrich:65906 | (1:300) |
| Peptide, recombinant protein | VEGF-165 | Peprotech | Peprotech:100–20 | |
| Sequence-based reagent | siRNA against CAMSAP2#1 | Jiang et al. (2014); PMID: 24486153 | 5’- GAATACTTCTTGACGAGTT-3' | |
| Sequence-based reagent | siRNA against CAMSAP2#2 | Jiang et al. (2014); PMID: 24486153 | 5’- GTACTGGATAAATAAGGTA-3’ | |
| Sequence-based reagent | siRNA against CAMSAP3 | Noordstra et al. (2016); PMID: 27802168 | 5’-GCATTCTGGAGGAAATTGA-3’ | |
| Sequence-based reagent | siRNA against AKAP450 | Hurtado et al. (2011); PMID: 21606206 | 5’-AUAUGAACACAGCUUAUGA-3’ | |
| Sequence-based reagent | siRNA against MMG | Roubin et al. (2013); PMID: 23430395 | 5’-AGAGCGAGATCATGACTTA-3’ | |
| Sequence-based reagent | siRNA against CPAP | Tang et al. (2009); PMID: 19503075 | 5’- AGAAUUAGCUCGAAUAGAA-3’ | |
| Sequence-based reagent | morpholino against CAMSAP2b (Danio rerio) | Genetools | ATACAGATGgcaagtcttttacatc | |
| Sequence-based reagent | primers for CAMSAP2b (Danio rerio) amplification | This paper | see sequences in the zebrafish experiment section | |
| Commercial assay or kit | AMAXA huvecs nucleofector kit | Lonza | Lonza:VPB-1002 | |
| Recombinant DNA reagent | pLenti-RhoA2G | Addgene | Addgene:40179 | |
| Recombinant DNA reagent | pLVIN-Rac1-bs Rac1 | Bouchet et al. (2016); PMID: 27939686 | ||
| Chemical compound, drug | rat tail collagen I | Corning | Corning:734–1085 | |
| Chemical compound, drug | Centrinone | Wong et al. (2015); PMID: 25931445 | ||
| Chemical compound, drug | Y27632 | Sigma-Aldrich | Sigma-Aldrich:Y0503 | |
| Chemical compound, drug | Blebbistatin | Enzo Life Science | Enzo Life Science:BML-EI315-0005 | |
| Software, algorithm | ImageJ SOS plugin | Yao et al., 2017; PMID: 28324611 | ||
| Software, algorithm | ImageJ radiality plugin | https://github.com/ekatrukha/radialitymap | Katrukha, 2017. radialitymap. Github. https://github.com/ekatrukha/radialitymap cf1e78f | |
| Software, algorithm | imageJ curve tracing plugin | https://github.com/jalmar/CurveTracing | Teeuw and Katrukha, 2015. CurveTracing. Github. https://github.com/jalmar/CurveTracing 960852 f |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.33864.050
