Antibodies |
ACTIN |
Sigma-Aldrich |
Cat# A2066 RRID:AB_476693 |
Alexa Donkey anti-Chicken IgG (H+L) 647 |
Thermo Fisher Scientific |
Cat# A78952 RRID:AB_2921074 |
Alexa Donkey anti-Goat IgG (H+L) 488 |
Thermo Fisher Scientific |
Cat# A11055 RRID:AB_2534102 |
Alexa Donkey anti-Goat IgG (H+L) 555 |
Thermo Fisher Scientific |
Cat# A21432 RRID:AB_2535853 |
Alexa Donkey anti-Mouse IgG (H+L) 488 |
Thermo Fisher Scientific |
Cat# A210202 RRID:AB_141607 |
Alexa Donkey anti-Mouse IgG (H+L) 555 |
Thermo Fisher Scientific |
Cat# A31570 RRID:AB_2536180 |
Alexa Donkey anti-Mouse IgG (H+L) 647 |
Thermo Fisher Scientific |
Cat# A31571 RRID:AB_162542 |
Alexa Donkey anti-Rabbit IgG (H+L) 488 |
Thermo Fisher Scientific |
Cat# A21206 RRID:AB_2535792 |
Alexa Donkey anti-Rabbit IgG (H+L) 555 |
Thermo Fisher Scientific |
Cat# A31572 RRID:AB_162543 |
ChAT |
Millipore |
Cat#AB144P RRID:AB_2079751 |
HRP Donkey anti-Goat IgG (H+L) Secondary Antibody |
Thermo Fisher Scientific |
Cat# PA1-28664 RRID:AB_10990162 |
HRP Goat anti-Mouse IgG (H+L) Secondary Antibody |
Thermo Fisher Scientific |
Cat# 31430 RRID:AB_228307 |
HRP Goat anti-Rabbit IgG (H+L) Secondary Antibody |
Thermo Fisher Scientific |
Cat# 31460 RRID:AB_228341 |
ITGB1 |
Millipore |
Cat#MAB2079Z RRID:AB_2233964 |
ITGB4 |
Millipore |
Cat#MAB1946 RRID:AB_2924963 |
FAK |
Cell Signaling |
Cat#3285S RRID:AB_2269034 |
FOXA2 |
Santa Cruz Biotechnology |
Cat#sc-101060 RRID:AB_1124660 |
GABA |
Sigma-Aldrich |
Cat#A2052 |
GFP |
Abcam |
Cat#ab6673 |
ILK |
Cell Signaling |
Cat#3862 |
ISL1/2 |
DSHB |
Cat# 39.4D5, RRID:AB_2314683 |
Ki67 |
Abcam |
Cat#ab66155 RRID:AB_1140752 |
Laminin alpha-1 |
Santa Cruz Biotechnology |
Cat#sc-74417 RRID:AB_1125087 |
MAP2 |
Biolegend |
Cat#840601 RRID:AB_2565455 |
NEUN |
Biolegend |
Cat#834502 RRID:AB_2734601 |
p-FAK |
Cell Signaling |
Cat#3281S RRID:AB_331079 |
PSD95 |
NeuroMab |
Cat#75028 RRID:AB_2292909 |
TUJ1 |
Biolegened |
Cat#802001 RRID:AB_2564645 |
SOD1 |
Proteintech |
Cat#10269-1-AP RRID:AB_2193750 |
SYN1 |
Millipore |
Cat#AB1543 RRID:AB_2200400 |
SYP |
Abcam |
Cat#ab32127 RRID:AB_2286949 |
Ubiquitin (UB) |
Santa Cruz Biotechnology |
Cat#sc-8017 RRID:AB_628423 |
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Bacterial and virus strains |
One Shot Stbl3 Chemically Competent E. coli
|
Thermo Fisher Scientific |
Cat# C737303 |
One Shot TOP10 Chemically Competent E. coli
|
Thermo Fisher Scientific |
Cat# C404010 |
HIV-1-based lentivirus |
This paper |
Zufferey et al., 1998 [123] |
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Biological samples |
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Chemicals, peptides, and recombinant proteins |
Accutase |
Innovative Cell Technologies |
Cat# AT 104-500 |
(3-Aminopropyl)triethoxysilane |
Sigma-Aldrich |
Cat# 440140 |
Ascorbic Acid |
Sigma-Aldrich |
Cat# A4403 |
B27 supplement |
Thermo Fisher Scientific |
Cat# 17504001 |
BDNF |
R&D Systems |
Cat# 248-BD |
CNTF |
R&D Systems |
Cat# 257NT/CF |
DAPI (4’,6-Diamidino-2-Phenylindole, Dihydrochloride) |
Thermo Fisher Scientific |
Cat# D1306 |
DAPT |
DNSK International |
Cat# 208255-80-5 |
Dimethylformamide |
Sigma-Aldrich |
Cat# RTC000098 |
1,6-Diphenyl-1,3,5-hexatriene, |
Sigma-Aldrich |
Cat# D208000 |
DMEM/F12 |
Thermo Fisher Scientific |
Cat# 11320-082 |
DNase (D2) |
Worthington |
Cat# LK003172
|
Doxycycline |
Sigma-Aldrich |
Cat# D9891-5G |
ECL (PierceTM ECL) substrate |
Thermo Fisher Scientific |
Cat# 32106 |
ECL (Radiance ECL) substrate |
Azure Biosystems |
Cat# AC2204 |
Epredia Immu-Mount |
Thermo Fisher Scientific |
Cat# 9990402 |
FBS - HyClone |
VWR |
Cat# 16777-006 |
Fibronectin |
Thermo Fisher Scientific |
Cat# PHE0023 |
GDNF |
R&D Systems |
Cat# 212-GD |
45% glucose solution |
Sigma-Aldrich |
Cat# G8769-100ML |
Glutamax |
Thermo Fisher Scientific |
Cat# 35050-061 |
Glutaraldehyde |
Fisher Scientific |
Cat# 50-262-18 |
Heparin Sulfate |
Sigma Aldrich |
Cat# H3149 |
Halt protease and phosphatase inhibitor cocktail |
Thermo Fisher Scientific |
Cat# 78447 |
IKVAV-PA1: C16VVAAEEEEGIKVAV |
Alvarez et al, 2021 [48]. |
N/A |
IKVAV-PA2: C16AAGGEEEEGIKVAV |
Alvarez et al, 2021 [48]. |
N/A |
IKVAV-PA3: C16VEVAAEEEEGIKVAV |
This paper |
N/A |
IKVAV peptide |
Olbrich et al, 1996 [48]. |
N/A |
Knockout DMEM |
Thermo Fisher Scientific |
Cat# 10829-018 |
Knockout Replacement Serum |
Thermo Fisher Scientific |
Cat# 10828-010 |
Laminin |
Life Technologies |
Cat# 23017-015 |
Laminin 211 |
BioLamina |
Cat# LN211-0501 |
Laminin 521 |
BioLamina |
Cat# LN521-05 |
LDN-193189 |
DNSK International |
Cat# 1062368-24-4 |
Matrigel |
Corning |
Cat# BD354277
|
2-Mercaptoethanol |
Thermo Fisher Scientific |
Cat# 21985-023 |
Micro-90 detergent |
Sigma-Aldrich |
Cat# Z281565 |
mTeSR1 media |
STEMCELL |
Cat# 85850 |
N2 supplement |
Thermo Fisher Scientific |
Cat# 17502001 |
NEAA |
Fisher Scientific |
Cat# 11140050 |
Neurobasal |
Thermo Fisher Scientific |
Cat# 21103-049 |
Normal Donkey Serum |
Jackson Immuno Research |
Cat# 017-000-001 |
Paraformaldehyde 16% |
Fisher Scientific |
Cat# 50-980-487 |
Polyethylenimine hydrochloride |
Sigma-Aldrich |
Cat# 764965 |
1X PBS (TC) |
Fisher Scientific |
Cat# MT21040CV |
Poly-D-Lysine hydrobromide |
Sigma-Aldrich |
Cat# P6407 |
Poly-DL-ornithine hydrobromide |
Sigma-Aldrich |
Cat# P0671 |
Pen/Strep |
Fisher Scientific |
Cat# 45000-650 |
Puromycin |
Sigma-Aldrich |
Cat# P8833 |
Retinoic Acid |
Sigma-Aldrich |
Cat# R2625 |
ROCK inhibitor (Y-27632) |
DNSK International |
Cat# 129830-38-2 |
SB431542 |
DNSK International |
Cat# 301836-41-9 |
Scr-PA1: C16VVAAEEEEGVVIAK |
Alvarez et al, 2021 [48]. |
N/A |
Scr-PA2: C16AAGGEEEEGVVIAK |
Alvarez et al, 2021 [48]. |
N/A |
Scr-PA3: C16VEVAAEEEEGVVIAK |
This paper |
N/A |
Smoothened-Agonist (SAG) |
DNSK International |
Cat# 364590-63-6 |
SU5402 |
DNSK International |
Cat# 215542-92-3 |
3-(trimethylsilyl) propionic-2,2,3,3-d4 acid, sodium salt |
Sigma-Aldrich |
Cat# 269913 |
Tetrahydrofuran (THF) |
Sigma-Aldrich |
Cat# 78445 |
TrypLE Express |
Thermo Fisher Scientific |
Cat# 12604-013 |
XAV939 |
Stemgent |
Cat# 04-00046 |
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Critical commercial assays |
BCA protein assay |
Thermo Fisher Scientific |
Cat# 23225 |
CytoTox 96 Non-Radioactive Cytotoxicity Assay |
Promega |
Cat# G1780 |
HilyMax |
Dojindo Molecular Technologies |
Cat# H357-10 |
Lipofectamine RNAiMAX |
Invitrogen |
Cat# 13778030 |
Superscript IV Reverse Transcriptase |
Invitrogen |
Cat# 18090010 |
iTaq Universal SYBR Green Supermix |
Bio-Rad |
Cat# 1725124 |
TRIzol Reagent |
Thermo Fisher Scientific |
Cat# 15596026 |
10plex Tandem Mass Tag System |
Thermo Fisher Scientific |
Cat# 90110 |
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Deposited data |
TMT LC-MS/MS data |
This paper |
Table S1
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Experimental models: Cell lines |
Human iPSC line 18a |
Boulting et al, 2011 [104]. Harvard University |
RRID:CVCL_8993 |
Human iPSC line 11a |
Boulting et al, 2011 [104]. Harvard University. |
RRID:CVCL_8987 |
Human iPSC SOD1+/4AV (39b) |
Kiskinis et al., 2014 [92]. Harvard University |
RRID:CVCL_X725 |
Human iPSC SOD1+/+ (39b-cor) |
Kiskinis et al., 2014 [92]. Harvard University |
N/A |
HEK293-FT |
ATCC |
RRID:CVCL_6911 |
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Experimental models: Organisms/strains |
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Oligonucleotides |
TUBB3: GGCCAAGGGTCACTACACG (Forward); GCAGTCGCAGTTTTCACACTC (Reverse). |
This paper |
N/A |
ISL1: AGATCAGCCTGCTTTTCAGC (Forward); TCATGCCTCAATAGGACTGG (Reverse) |
This paper |
N/A |
MNX1: GAACACCAGTTCAAGCTCAACA (Forward); GCTGCGTTTCCATTTCATTCG (Reverse). |
This paper |
N/A |
LHX3: GGAGAGCGTTTACTGCAAGGA (Forward); TTGGCGGTTTCGTAGTCCG (Reverse). |
This paper |
N/A |
FOXA2: GGAGCAGCTACTATGCAGAGC (Forward); CGTGTTCATGCCGTTCATCC (Reverse). |
This paper |
N/A |
GPI: GTGTACCTTCTAGTCCCGCC (Forward); GGTCAAGCTGAAGTGGTTGAAGC (Reverse). |
This paper |
N/A |
RPL0: TCTACAACCCTGAAGTGCTTGAT (Forward); CAATCTGCAGACAGACACTGG (Reverse). |
This paper |
N/A |
siITB1 (siRNA ID:S7576) |
Thermo Fisher Scientific |
Cat# 4390824 |
siScramble |
Thermo Fisher Scientific |
Cat# 4390843 |
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Recombinant DNA |
FUW-M2rtTA |
Addgene |
RRID:Addgene_20342 |
Lentiviral GFP plasmid |
This paper / VectorBuilder |
VB181102-1195sys |
Lentiviral ITGB1::GFP plasmid |
This paper / VectorBuilder |
VB211104-1470afj |
pMD2.G |
Addgene |
RRID:Addgene_12259 |
psPAX2 |
Addgene |
RRID:Addgene_12260 |
pTet-O-Ngn2-Puro |
Addgene |
RRID:Addgene_52047 |
Tet-O-FUW-EGFP |
Addgene |
RRID:Addgene_30130 |
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Software and algorithms |
Adobe Photoshop 7.0 |
Adobe Systems Incorporated |
RRID:SCR_014199 |
Avogadro |
Open Molecules |
RRID:SCR_015983 |
Axion’s Integrated Studio (AxIS) software |
Axion Biosystems |
RRID:SCR_016308 |
BioRender |
BioRender |
RRID:SCR_018361 |
pCLAMP 11.1 (Clampex 11 software) |
Molecular devices |
RRID:SCR_011323 |
Crystallography Data Analysis Software |
Toby and Von Dreele, 2013 [124]. |
https://subversion.xray.aps.anl.gov/trac/pyGSAS
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Cytoscape / Metascape |
Zhou et al., 2019 [125]. |
RRID:SCR_016620 |
Comparative GO |
Fruzangohar et al., 2017 [126]. |
https://www.comparativego.com
|
Database for Annotation, Visualization and Integrated Discovery (DAVID) |
Huang et al., 2009 [127]. |
RRID:SCR_001881 |
Easy Electrophysiology |
Easy Electrophysiology Ltd |
RRID:SCR_021190 |
Fiji |
Schindelin et al., 2012 [128]. |
RRID:SCR_002285 |
GROMACS 5.0.4 Software |
GROMACS development team |
https://doi.org/10.5281/zenodo.7323393.
|
Imaris |
Oxford Instruments Group |
RRID:SCR_007370 |
Martinize.py, 2.0 |
Martini - General Purpose Coarse-Grained Force Field |
http://cgmartini.nl/images/tools/martinize/martinize-2.6/martinize.py
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MATLAB |
The MathWorks, Inc. |
RRID:SCR_001622 |
Mx™ Software |
Zygo Corporation |
https://www.zygo.com
|
NanoScope analysis software |
Bruker |
http://nanoscaleworld.bruker-axs.com/nanoscaleworld/media/p/775.aspx
|
NIS-Elements Advanced Research version 5 |
Nikon Instruments Inc. |
RRID:SCR_002776 |
Prism7 |
GraphPad Software |
RRID:SCR_002798 |
Protein unfolding nano-indentation analysis software |
Punias |
punias@free.fr
|
Scaffold v5 |
Proteome Software Inc. |
RRID:SCR_014345 |
Small Angle Scattering Analysis Software |
SasView |
https://www.sasview.org
|
VMD-Visual Molecular Dynamics |
University of Illinois at Urbana-Champaign |
https://www.ks.uiuc.edu/Research/vmd/
|
WinFluor Software |
University of Strathclyde |
http://spider.science.strath.ac.uk/sipbs/software_winfluor.htm
|
Cinema 4D version R20 |
Maxon Computer GMBH |
https://www.maxon.net/es/downloads/cinema-4d-past-versions
|
Other |
CD-1 IGS (Crl:CD1(ICR)) mice for primary glia cultures and spinal cord tissue |
Charles River |
Cat#22; RRID:IMSR_CRL:022 |
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