Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Rattus norvegicus) |
Wistar | SLC | RRID:RGD_2314928 | |
| Gene (Rattus norvegicus) |
Wistar | CLEA Japan | RRID:RGD_12879431 | |
| Gene (Mus musculus) |
C57BL/6 | SLC | RRID:MGI:5658686; RRID:MGI:5295404 |
|
| Gene (Mus musculus) |
C57BL/6 | CLEA Japan | RRID:MGI:5658686; RRID:MGI:2160139 |
|
| Genetic reagent (Mus musculus) |
Shootin1 gene knockout |
This paper | Please see 'Generation of Shootin1 knockout mice' in Materials and methods section |
|
| Cell line (Homo sapiens) |
HEK293T cell | ATCC | Cat# CRL_3216; RRID:CVCL_0063 |
|
| Antibody | anti-shootin1a peptide sequence (rabbit polyclonal) |
This paper | Rabbit polyclonal; against aa 450–456; Immunohistochemistry: (1:5000) Immunoblot: (1:5000) |
|
| Antibody | anti-shootin1 antibody (rabbit polyclonal) |
PMID: 17030985 (Toriyama et al., 2006); PMID: 23453953 (Toriyama et al., 2013) |
Immunoblot: (1:1000) | |
| Antibody | anti-pSer101-shootin1 antibody (rabbit polyclonal) |
PMID: 23453953 (Toriyama et al., 2013) | Immunoblot: (1:1000) | |
| Antibody | anti-pSer249-shootin1 antibody (rabbit polyclonal) |
PMID: 23453953 (Toriyama et al., 2013) | Immunoblot: (1:5000); Immunofluorescence(1:1000) |
|
| Antibody | anti-NCAM-L1 (C-20) antibody (goat polyclonal) |
Santa Cruz Biotechnology | Cat# sc-1508; RRID:AB_631086 | Immunoblot: (1:2000); Immunofluorescence: (1:1000); Immunohistochemistry: (1:1000) |
| Antibody | anti-Neurofilament antibody 2H3 (mouse monoclonal) |
DSHB | Cat# 2H3; RRID:AB_531793 | Immunohistochemistry: (1:2000) |
| Antibody | anti-TAG-1 antibody 4D7 (mouse monoclonal) |
DSHB | Cat# 4D7/TAG1; RRID:AB_531775 | Immunohistochemistry: (1:100) |
| Antibody | anti-FLAG(DDDDK) tag antibody (rabbit polyclonal) |
MBL | Cat# PM020; RRID:AB_591224 | Immunoblot: (1:1000) |
| Antibody | anti-Myc tag antibody (rabbit polyclonal) |
MBL | Cat# 562–5; RRID:AB_591116 | Immunoblot: (1:2000) |
| Antibody | anti-GST tag antibody (goat polyclonal) |
GE Healthcare | Cat# 27-4577-01; RRID:AB_771432 | Immunoblot: (1:3000) |
| Antibody | anti-His tag antibody (goat polyclonal) |
Wako | Cat# 014–23221 | Immunofluorescence (1:500) |
| Antibody | anti-rabbit IgG secondary antibody, Alexa Fluor 594 (from donkey) |
Jackson immune research | Cat# 711-585-152; RRID:AB_2340621 |
Immunofluorescence: (1:1000); Immunohistochemistry: (1:1000) |
| Antibody | anti-goat IgG secondary antibody, Alexa Fluor 488 (from donkey) |
Invitrogen, Thermo Fisher Scientific |
Cat# A-11055; RRID:AB_2534102 |
Immunofluorescence: (1:1000); Immunohistochemistry: (1:1000) |
| Antibody | anti-mouse IgG secondary antibody, Alexa Fluor 488 (from goat) |
Invitrogen, Thermo Fisher Scientific |
Cat# A-11029; RRID:AB_2534088 |
Immunofluorescence: (1:1000) |
| Antibody | anti-rabbit IgG, Whole Ab ECL antibody, HRP Conjugated (from donkey) |
GE Healthcare | Cat# NA934; RRID:AB_772206 |
Immunoblot: (1:2000) |
| Antibody | anti-goat IgG, HRP conjugate, Species Adsorbed: H, M, R, Ch, Gp, Eq, Ht, Rb antibody (from donkey) |
Millipore | Cat# AP180P; RRID:AB_92573 |
Immunoblot: (1:2000) |
| Recombinant DNA reagent |
pCMV-myc vector | Stratagene, Agilent | Cat# 211173 | |
| Recombinant DNA reagent |
pCMV-Flag vector | Strategene, Agilent | Cat# 211172 | |
| Recombinant DNA reagent |
pCAGGS vector | PMID: 1660837 (Niwa et al., 1991) Strategene, Addgene |
Collection number LMBP2453 |
This vector was provided by J. Miyazaki, Osaka University, Osaka, Japan; Niwa et al. (1991) (PMID: 1660837) |
| Recombinant DNA reagent |
pGEX-6P-1 | GE Healthcare | Cat# 28954648 | |
| Recombinant DNA reagent |
pCMV-mRFP-actin vector | PMID: 29483251 (Abe et al., 2018) |
||
| Recombinant DNA reagent |
pCMV-myc-shootin1a vector | PMID: 17030985 (Toriyama et al., 2006) |
||
| Recombinant DNA reagent |
pCMV-Flag-L1-CAM-ICD (intracellular domain) vector |
This paper | ||
| Recombinant DNA reagent |
pCMV-dominant negative myc-Pak1vector |
PMID: 26261183 (Kubo et al., 2015) |
||
| Recombinant DNA reagent |
pCMV-constitutively active myc-Pak1 vector |
PMID: 26261183 (Kubo et al., 2015) |
||
| Recombinant DNA reagent |
pCAGGS-myc | PMID: 17030985 (Toriyama et al., 2006) |
||
| Recombinant DNA reagent |
pCAGGS-myc-GST vector | PMID: 17030985 (Toriyama et al., 2006) |
||
| Recombinant DNA reagent |
pCAGGS-myc- shootin1a (1-125) vector |
This paper | ||
| Recombinant DNA reagent |
pCAGGS-EGFP | This paper | ||
| Recombinant DNA reagent |
pCAGGS-EGFP- shootin1a(1-125) |
This paper | ||
| Recombinant DNA reagent |
pGEX-shootin1a-WT vector | PMID: 26261183 (Kubo et al., 2015) |
||
| Recombinant DNA reagent |
pGEX-shootin1a-DD (phopho-mimic shootin1a) vector |
PMID: 26261183 (Kubo et al., 2015) |
||
| Recombinant DNA reagent |
pGEX-myc-shootin1a-DD (phopho-mimic shootin1a) vector |
PMID: 26261183 (Kubo et al., 2015) |
||
| Recombinant DNA reagent |
pGEX-myc-shootin1a- (1-125) vector |
PMID: 26261183 (Kubo et al., 2015) |
||
| Recombinant DNA reagent |
pGEX-myc-shootin1a- (125-260) vector |
PMID: 26261183 (Kubo et al., 2015) |
||
| Recombinant DNA reagent |
pGEX-myc-shootin1a- (217-456) vector |
PMID: 26261183 (Kubo et al., 2015) |
||
| Recombinant DNA reagent |
pGEX-myc-shootin1a- (261-377) vector |
PMID: 26261183 (Kubo et al., 2015) |
||
| Recombinant DNA reagent |
pGEX-L1-CAM-ICD (intracellulardomain) vector |
This paper | ||
| Recombinant DNA reagent |
RNAi shootin1a vector (miRNA) |
PMID: 17030985 (Toriyama et al., 2006); PMID: 23453953 (Toriyama et al., 2013) |
||
| Recombinant DNA reagent |
RNAi-refractory shootin1a-WT vector |
PMID: 23453953 (Toriyama et al., 2013) |
||
| Recombinant DNA reagent |
RNAi-refractory shootin1a-DD (phopho- mimic shootin1a) vector |
PMID: 23453953 (Toriyama et al., 2013) |
||
| Peptide, recombinant protein |
recombinant Netrin-1 protein (from mouse) |
R and D systems | Cat# 1109-N1-025 | No CF (No carrier protein free) |
| Peptide, recombinant protein |
FLAG peptide | Sigma-Aldrich | Cat# F3290 | |
| Peptide, recombinant protein |
Laminin Solution, from Mouse EHS Tumor |
Wako | Cat# 120–05751 | |
| Peptide, recombinant protein |
L1-CAM-Fc | PMID: 18519736 (Shimada et al., 2008) |
N/A | |
| Peptide, recombinant protein |
Prescission protease | GE Healthcare | Cat# 27084301 | |
| Peptide, recombinant protein |
Recombinant Pak1 | Life technologies, Thermo Fisher Scientific |
Cat# PV3820 | |
| Commercial assay or kit |
Rat Neuron Nucleofector kits (25 RCT) |
Lonza | Cat# VPG-1003 | |
| Chemical compound, drug |
anti-FLAG M2 antibody affinity gel (mouse monoclonal) |
Sigma-Aldrich | Cat# A2220; RRID:AB_10063035 |
|
| Chemical compound, drug |
7-amino-4- chloromethylcoumarin (CMAC) |
Invitrogen, Thermo Fisher Scientific |
Cat# C2110 | |
| Chemical compound, drug |
4,6-diamidino-2- phenylindole(DAPI) |
Roche | Cat# 10236276001 Roche |
DAPI stain (1:1000) |
| Chemical compound, drug |
1,1'-dioctadecyl- 3,3,3',3'-tetramethyl- indocarbocyanine dye (DiI) |
Invitrogen, Thermo Fisher Scientific |
Cat# D3911 | |
| Chemical compound, drug |
Glutathione sepharose 4B | GE Healthcare | Cat# 17-0756-01 | |
| Chemical compound, drug |
Protein G-sepharose 4B | GE Healthcare | Cat# 6511–5 | |
| Chemical compound, drug |
Polydimethylsiloxane (PDMS) | Dow Corning Toray, Japan | Cat# 3255981 | |
| Chemical compound, drug |
Silicone oil (Barrier coat No.6) | ShinEtsu, Japan | Cat# 06003 | |
| Chemical compound, drug |
PhosSTOP | Roche | Cat# 4906845001 | |
| Software, algorithm | Image J | https://imagej.nih.gov/ij/ | RRID:SCR_003070 | |
| Software, algorithm | Fiji | http://fiji.sc | RRID:SCR_002285 | |
| Software, algorithm | Graphpad prism 7 | Graphpad software | RRID:SCR_002798 | |
| Software, algorithm | R Project for Statistical Computing |
http://www.r-project.org/ | RRID:SCR_001905 | |
| Software, algorithm | Matlab | http://www.mathworks.com/products/matlab/ | RRID:SCR_001622 | |
| Software, algorithm | Microsoft Excel 2016 | Microsoft https://www.microsoft.com |
||
| Other | Amicon ultra-4 centrifugal filter devices |
Millipore | Cat# UFC800324 |