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Table 1 Key resources used in this paper

From: A synthetic pregnenolone analog promotes microtubule dynamics and neural development

Antibodies

Source

Identifier

Note

Mouse monoclonal anti-acetylated tubulin, clone 6-11B-1

Sigma

Cat#T6793

1:500

Rabbit monoclonal anti-III-beta tubulin [EP1569Y]

Abcam

Cat#ab52623

1:500

Rabbit polyclonal anti-beta tubulin

Proteintech

10094-1-AP

1:200

Goat anti-mouse IgG (H+L) superclonal recombinant secondary antibody, Alexa Fluor 555

Invitrogen

A28180

1:1000

Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Oregon Green 488

Invitrogen

O-11038

1:4000

Experimental models

Source

Identifier

Note

Mouse: C57BL/6J strain

National Applied Research Laboratories

  

Zebrafish: TL strain

   

COS1 cells

   

Plasmids

Source

Identifier

Note

pSC2 + mOrange2-Flag-CLIP1

This paper

N/A

Fluorescent CLIP1 for comet tracking

Steroids

Source

Identifier

Note

Pregnenolone, 5-pregnen-3β-ol-20-one

Sigma-Aldrich

P9129

 

P5 derivatives

This paper

N/A

 

Reagents for cell culture

Source

Identifier

Note

Dulbecco’s Modified Eagle Medium

Gibco

12100-046

COS1 culture

HyClone Characterized Fetal Bovine Serum

GE Life Sciences

SH30396.03

COS1 culture

Neurobasal Plus Medium

Gibco

A35829-01

Primary neuronal culture

Glutamax-I (100X)

Gibco

35050-061

Primary neuronal culture

B27 Supplement (50X)

Gibco

17504044

Primary neuronal culture

Penicillin–Streptomycin, liquid

Gibco

15140-122

Primary neuronal culture

HEPES, 1 M buffer solution

Gibco

15630-080

Primary neuronal culture

HBSS, 10X

Gibco

14185-052

Primary neuronal culture

Percoll pH 8.5–9.5

Sigma-Aldrich

P1644

Primary neuronal culture

Poly-L-ornithine, 0.01%

Sigma-Aldrich

P4957

Primary neuronal culture

Trypsin–EDTA

Gibco

25300-054

Primary neuronal culture

Trypsin inhibitor from Glycine max (soybean)

Sigma-Aldrich

T6522

Primary neuronal culture

DNase I

Roche

04716728001

Primary neuron harvest

In vitro microtubule polymerization assay

Source

Identifier

Note

Tubulin protein (> 99% pure): porcine brain

Cytoskeleton

T240

 

Tubulin protein (rhodamine): porcine brain

Cytoskeleton

TL590M

 

GpCpp

Jena Biosciences

NU-405

 

Paclitaxel from Taxus brevifolia, ≥ 95% (HPLC), powder

Sigma Aldrich

T7402

 

0.5 mL, Open-Top Thickwall Polycarbonate Tube, 8 × 34 mm

Beckman Coulter

343776

 

Device

Source

Identifier

Note

Neon Transfection System

Invitrogen

MPK5000

 

DeltaVision Core

GE Healthcare

N/A

Live cell imaging for comet tracking

Zeiss LSM710 confocal microscope

Carl Zeiss

N/A

Neurons imaging

Software

Source

Note

 

ImarisFileConverter × 64 9.7.2

Oxford Instruments

  

Imaris × 64 9.7.2

Oxford Instruments

Quantifying cerebellum volume

 

SoftMax Pro 5.3

Molecular Devices

  

EnSpire

PerkinElmer

  

ImageJ/Fiji

https://imagej.net/Fiji

Base program for MTrackJ

 

softWoRx

Applied Precision

  

MTrackJ

[47]

CLIP1 comet quantification

 

ZEN 2.6

Carl Zeiss

Image processing

 

GraphPad Prism 8.0.0

http://www.graphpad.com

Statistical analysis

 

Reagents for transfection and staining

Source

Identifier

Note

Neon Transfection System 100 µL Kit

Invitrogen

MPK10096

pSC2+mOrange2-Flag-CLIP1 transfection

DAPI

Life Technologies

D1306

1:10000

Rhodamine phalloidin

Cytoskeleton

PHDR1

 

ProLong Gold Antifade Reagent

Invitrogen

P36930

 
  1. N/A, not applicable