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. 2022 Jun 7;25(7):104542. doi: 10.1016/j.isci.2022.104542
REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies

Mouse Anti-Villin1 Abcam Cat#ab3304; RRID: AB_303688
Rabbit Anti-E-cadherin Cell Signaling Technology Cat#3195; RRID: AB_2291471
Rabbit Anti-SGLT1 Merck Cat#07-1417
Goat Anti-Apolipoprotein B-100 Abcam Cat#ab7616; RRID: AB_305987
Cy3-conjugated donkey anti-mouse IgG Jackson ImmunoResearch Cat#711-166-150
Cy3-conjugated donkey anti-rabbit IgG Jackson ImmunoResearch Cat#711-166-152
Alexa Fluor 488-conjugated donkey anti-rabbit IgG Jackson ImmunoResearch Cat#711-546-152
HRP-conjugated donkey anti-goat IgG Jackson ImmunoResearch Cat#705-035-147

Bacterial and virus strains

One Shot™ Stbl3™ Chemically Competent E. coli Thermo Fisher Scientific C737303
Mix & Go! Competent Cells-TG1 Zymo Research T3017

Chemicals, peptides, and recombinant proteins

DMEM, high glucose FUJIFILM Wako 044-29765
E-MEM with L-Glutamine and Phenol Red FUJIFILM Wako 051-07615
Advanced DMEM/F12 Thermo Fisher Scientific 12634-028
MEM Non-Essential Amino Acids Solution (100X) Thermo Fisher Scientific 11140050
Fetal bovine serum Thermo Fisher Scientific 10270-106
GlutaMAX Supplement (100X) Thermo Fisher Scientific 35050061
HEPES (1 M) Thermo Fisher Scientific 15630080
30% Bovine serum albumin FUJIFILM Wako 015-23871
Matrigel Corning 354234
Y-27632 FUJIFILM Wako 257-00511
SB202190 Sigma-Aldrich S7067
CultureSure A83-01 FUJIFILM Wako 039-24111
Gentamicin Sulfate Solution (50 mg/mL) Nacalai Tesque 11980-14
Hexadimethrine bromide (Polybrene) Sigma-Aldrich 107689
Rifampicin FUJIFILM Wako 189-01001
Oleic acid Nacalai Tesque 25630-51
CP-346086 dihydrate Sigma-Aldrich PZ0103
α-methyl-D-glucopyranoside (AMG) Tokyo Chemical Industry M0228
[14C] AMG PerkinElmer NEC659
N-(4-hydroxyphenyl)retinamide (4-HPR) Tokyo Chemical Industry H1464
Wnt3a, R-spondin1, Noggin CM Takahashi et al., 2018 N/A
Lipofectamine 3000 Transfection Reagent Thermo Fisher Scientific L300015
Lipofectamine Stem Transfection Reagent Thermo Fisher Scientific STEM00015
TrypLE Express Thermo Fisher Scientific 12604013
Cell Recovery Solution BD Biosciences 354253
Recombinant mouse EGF PeproTech 315-09
Recombinant human HGF R&D systems 294-HG-005
Tissue-Tek Optimal Cutting Temperature compound Sakura Finetechnical 4583
Fluoromount Diagnostic BioSystems K024
Prestained XL-Ladder Broad Intégrale SP-2120
30% (w/v)-Acrylamide-bis (29:1) Nacalai Tesque 06141-35
DAPI Fluorescence Stain, 1000X Cell Biolabs 112002
Immobilon-P Merck IPVH00010
Blocking One Nacalai Tesque 03953-95

Critical commercial assays

SmGM-2 Bullet Kit Lonza CC-3182
ISOGEN NIPPON GENE 319-90211
RNeasy Mini Kit QIAGEN 74104
RNase free DNase set QIAGEN 79254
High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific 4368814
FASTStart Universal SYBR Green Master (ROX) Roche 4913914001
TaqMan Gene Expression Master Mix Thermo Fisher Scientific 4369016
QIAGEN Plasmid Midi Kit QIAGEN 12145
BD Cytofix/Cytoperm Fixation/Permeabilization Solution Kit BD Biosciences 554714
ECL Western Blotting Detection Reagents Cytiva RPN2106
Immobilon Western Chemiluminescent HRP Substrate Merck WBKLS0500
Nano-Glo Luciferase Assay System Promega N1130
HS RNA Kit Agilent DNF-472-0500
MGIEasy RNA Directional Library Prep Set MGI Tech 1000006386
Dynabeads mRNA Purification Kit Thermo Fisher Scientific 61006
dsDNA HS Assay Kit Thermo Fisher Scientific Q32851
DNBSEQ-G400RS High-throughput Sequencing Kit MGI Tech 1000016941
CellTiter-Glo 3D Cell Viability Assay Promega G9682

Deposited data

RNA-seq (primary-hIECs, Organoid-IECs, Differ-Caco-2) This paper DRA accession number: DRA013757

Experimental models: Cell lines

Mouse: L cells ATCC CRL-2648
Human: Caco-2 cells ATCC HTB-37
InEpC – Human Intestinal Epithelial Cells Lonza CC-2931
TkDN4-M Stem Cell Bank (The University of Tokyo) N/A
Human iPS cell-derived intestinal organoids Takahashi et al. (2018) N/A

Oligonucleotides

See Table S2 for information on qPCR probes and primers.

Recombinant DNA

p3xFLAG-CMV-10 Sigma-Aldrich E7658
p3xFLAG-CMV-10-hSGLT1 This paper N/A
pNL1.2 Promega N1011
pNL1.2-hCYP3A4 This paper N/A
pSV40-β-gal Promega E1081
pCAG-HIVgp RIKEN BioResource Research Center N/A
pCMV-VSV-G-RSV-Rev RIKEN BioResource Research Center N/A
CS-UbC-hSox2-IRES2-Venus RIKEN BioResource Research Center N/A
CS-hCYP3A4-Nluc This paper N/A

Software and algorithms

GraphPad Prism GraphPad Software https://www.graphpad.com/scientific-software/prism/
Cutadapt (v1.9.1) Martin (2011) https://github.com/marcelm/cutadapt
Sickle (v1.33) Joshi and Fass, 2011 https://github.com/najoshi/sickle
Hisat2 (v2.2.1) Kim et al. (2019) http://daehwankimlab.github.io/hisat2/
Samtools (v1.11) Li et al. (2009) http://www.htslib.org/
featureCounts (v2.0.0) Liao et al. (2014) https://www.rdocumentation.org/packages/Rsubread/versions/1.22.2/topics/featureCounts
R R Core Team 2020 https://www.r-project.org/