KEY RESOURCES TABLE
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Rabbit anti-mouse β-Actin antibody | Cell Signaling Technology | Cat#4967; RRID: AB_330288 |
Mouse anti-mouse AGXT antibody | Santa Cruz Biotechnology | Cat# SC-517388 |
Rabbit anti-mouse CYP7B1 antibody | Proteintech | CAT# 24889–1-AP RRID: AB_2879780 |
Mouse anti-mouse B-ACTIN antibody | Cell Signaling Technology | CAT#3700 RRID: AB_2242334 |
Rat anti-Galectin 3 antibody | Thermo Fisher Scientific | Cat#13-5301-81; RRID: AB_837112 |
CCL5 (RANTES) Recombinant Polyclonal Antibody | ThermoFisher Scientific | Cat# 710001, RRID: AB_2532515 |
800CW Donkey anti-mouse IRDye | LI-COR Biosciences | Cat# 926–32212; RRID: AB_621847 |
680RD Donkey anti-rabbit IRDye | LI-COR Biosciences | Cat# 926–68073; RRID: AB_10954442 |
800CW Donkey anti-rabbit IRDye | LI-COR Biotechnology | CAT# 926–32213 RRID: AB_621848 |
680RD Donkey anti-mouse IRDye | LI-COR Biotechnology | CAT# 926–68072 RRID: AB_10953628 |
Goat anti-Rat IgG (H+L) Secondary Antibody, HRP | ThermoFisher Scientific | Cat# 31470, RRID: AB_228356 |
Alexa Fluor 647 donkey anti-rat | Jackson ImmunoResearch Laboratories | Cat# 712-605-153; RRID: AB_2340694 |
Alexa Fluor 594 donkey anti-rabbit | Jackson ImmunoResearch Laboratories | Cat#711-585-152, RRID: AB_2340621 |
Bacterial and Virus Strains | ||
Stable Competent E. coli | New England Biolabs | Cat # C3040H |
AAV-TBG-GFP | This paper | N/A |
AAV-TBG-AGXT | This paper | N/A |
Biological Samples | ||
Serum from patients with or without sCAD | This paper | N/A |
Chemicals, Peptides, and Recombinant Proteins | ||
Polyethylenimine | Sigma-Aldrich | Cat# 408727 |
Dihydroethidium | Sigma-Aldrich | Cat# D7008 |
MitoTEMPO | Sigma-Aldrich | Cat# SML0737 |
Sodium oxalate | Sigma-Aldrich | Cat# 223433 |
Oil red O | Sigma-Aldrich | Cat# O0625 |
Sodium azide | Sigma-Aldrich | CAT#S2002 |
Proteinase K | Sigma-Aldrich | CAT# 1245680500 |
Trans-4-Hydroxy-L-proline | ACROS Organics | CAT#121785000 |
UltraPure™ Phenol:Chloroform:Isoamyl Alcohol | ThermoFisher Scientific | CAT#15593–031 |
Poloxamer 188 | Sigma-Aldrich | Cat# P5556 |
Harris hematoxylin | ThermoFisher Scientific | Cat# 842 |
Eosin Y | ThermoFisher Scientific | Cat# 832 |
Clarifier Bluing reagent | ThermoFisher Scientific | Cat# 7401 |
Bluing reagent | ThermoFisher Scientific | Cat# 7301 |
MitoSOX Red | ThermoFisher Scientific | Cat# M36008 |
Hoechst | ThermoFisher Scientific | Cat# H3570 |
Trizol reagent | ThermoFisher Scientific | Cat# 5596026 |
Canonical amino acid mix unlabeled standard | Cambridge Isotopes Laboratory | Cat# MSK-CAA-US-1 |
Canonical amino acid mix | Cambridge Isotopes Laboratory | Cat# MSK-CAA-1 |
O.C.T. Compound | Tissue-Tek | Cat# 4583 |
iQ SYBR Green Supermix | Bio-Rad | Cat# 1708880 |
Critical Commercial Assays | ||
Wako Cholesterol E | FUJIFILM medical system | Cat# 999–02601 |
RNeasy Maxi kit | QIAGEN | Cat# 75162 |
EndoFree Plasmid Maxi Kit | QIAGEN | Cat# 12362 |
Oxalate assay kit | Abcam | Cat# ab196990 |
Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit | R&D Systems | Cat# MJE00B |
Mouse/Rat CCL5/RANTES Quantikine ELISA Kit | R&D Systems | Cat# MMR00 |
Glucose test strips | Contour Next | Cat# NDC: 0193-7308-50 |
Seahorse XFp Cell Mito Stress Test Kit | Agilent Technologies | Cat# 103010–100 |
Gibson assembly kit | New England Biolabs | Cat# E5510 |
NEBNext Ultra II Directional RNA Library Prep Kit for Illumina | New England Biolabs | CAT# E7760L |
Poly(A) mRNA Magnetic Isolation Module | New England Biolabs | Cat# E7490L |
NEBNext Multiplex Oligos for Illumina Unique dual | New England Biolabs | Cat# E6440L |
Protease inhibitor | Thermo Fisher Scientific | Cat# A32965 |
Phosphatase inhibitor | Roche Diagnostics | Cat# 04906845001 |
SuperScript™ III First-Strand Synthesis System | ThermoFisher Scientific | Cat# 18080051 |
CCK8 assay kit | Sigma-Aldrich | Cat# 96992 |
SignalStain® DAB Substrate Kit | Cell Signaling Techonology | CAT#8059 |
Total Bile Acid Assays Kit | Cell Biolabs, INC | CAT#STA-631 |
Cholesterol fluorometric assay kit | Cayman Chemical | CAT#10007640 |
LabAssay™ Triglyceride kit | FUJIFILM medical system | CAT# 290–63701 |
Deposited Data | ||
RNA-sequencing | This paper | NCBI’s SRA (accession number: PRJNA671451) |
Raw data | This paper | Mendeley Data (doi:10.17632/n8nkdgd258.1) |
Experimental Models: Cell Lines | ||
HEK293T | ATCC | CRL-3216; RRID: CVCL_0063 |
L929 cells | ATCC | CRL-1; RRID: CVCL_0462 |
Bone marrow derived-macrophages | This paper | N/A |
Experimental Models: Organisms/Strains | ||
Mouse: C57BL/6J | The Jackson Laboratory | JAX: 000664; RRID: IMSR_JAX:000664 |
Mouse: Apoe−/− | The Jackson Laboratory | JAX: 002052; RRID: IMSR_JAX:002052 |
Mouse: Agxt−/− | Rom et al., 2020 | N/A |
Mouse: Agxt+/+/Apoe−/− | This paper | N/A |
Mouse: Agxt−/−/Apoe−/− | This paper | N/A |
Oligonucleotides | ||
Guide-RNA targeting the mouse Agxt exon1: 5’-GGGTCCGGGGCCCTCCAACC-3’ | Integrated DNA Technologies | N/A |
Primer sequences for Agxt genotyping: forward: 5’-ACACCTCCACTGTCCTGTCC-3’ | Integrated DNA Technologies | N/A |
Primer sequences for Agxt genotyping: reverse: 5’-GGTCAGATCTGCCTGCTACC-3’ | Integrated DNA Technologies | N/A |
Primer for Sanger sequencing: 5’-GCAGAGCTAGCTGGGAAATG-3’ | Integrated DNA Technologies | N/A |
Primer sequences for Apoe genotyping: forward: 5′-GCCTAGCCGAGGGAGAGCCG-3′ | Integrated DNA Technologies | N/A |
Primer sequences for Apoe genotyping: reverse 1: 5′-TGTGACTTGGGAGCTCTGCAGC-3′ | Integrated DNA Technologies | N/A |
Primer sequences for Apoe genotyping: reverse 2: 5′-GCCGCCCCGACTGCATCT-3’ | Integrated DNA Technologies | N/A |
Primer sequences for qPCR, see Table S2 | Integrated DNA Technologies | N/A |
Recombinant DNA | ||
pAdDeltaF6 | Jiandie Lin’s lab, University of Michigan; Guo et al., 2017 | N/A |
pAAV2/8 | Jiandie Lin’s lab, University of Michigan; Guo et al., 2017 | N/A |
pAAV-TBG-GFP | Jiandie Lin’s lab, University of Michigan; Guo et al., 2017 | N/A |
pAAV-TBG-MCS | Jiandie Lin’s lab, University of Michigan; Guo et al., 2017 | N/A |
AGXT Human Tagged ORF Clone | Origene | CAT# RG212899 |
Software and Algorithms | ||
ImageJ | National Institutes of Health | RRID:SCR_003070 https://imagej.nih.gov/ij/download.html |
GraphPad Prism | GraphPad Software | Version 8.0 RRID:SCR_002798 https://www.graphpad.com/scientific-software/prism/ |
R statistics | Hosted by Vienna, University of Economics and Business | http://www.R-project.org/. |
RStudio | RStudio Team (2020). | http://www.rstudio.com/.RRID:SCR_000432 |
IBM SPSS software | IBM Corporation | Version 23.0; RRID:SCR_019096 https://www.ibm.com/analytics/spss-statistics-software |
FastQC | Babraham Bioinformatics | Version 0.11.8 RRID:SCR_014583 https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ |
Trimmomatic | Bolger et al., 2014 | Version 0.35 RRID:SCR_011848 http://www.usadellab.org/cms/?page=trimmomatic |
HISAT2 | Kim et al., 2015 | Version 2.1.0.13; RRID:SCR_015530 http://daehwankimlab.github.io/hisat2/download/ |
HTSeq | Anders et al., 2015 | Version 0.6.0; RRID:SCR_005514 https://htseq.readthedocs.io/en/master/install.html |
Analyst software | Applied Biosystems | Version 1.6.2; RRID:SCR_015785 https://sciex.com/products/software/analyst-software |
LI-COR Image studio | LI-COR Biosciences | Version 2.1; RRID:SCR_015795 https://www.licor.com/bio/image-studio-lite/download |
SPOT Basic software | SPOT imaging | RRID:SCR_014313 https://www.spotimaging.com/software/spot-basic/ |
Other | ||
Mouse standard diet | LabDiet | Cat# 5L0D |
Mouse Western diet | Envigo | Cat# TD.88137 |
Mouse Western diet supplemented with hydroxy-L-proline (HLP) | Envigo | CAT# TD.190842 |
Dulbecco’s Modified Eagle Medium | Gibco | Cat# 11965092 |
Iscove’s Modified Dulbecco’s Media | Gibco | Cat# 12440053 |
Agilent Poroshell 120 EC-C18 column | Agilent Technologies | Cat# 699775–902 |
Superose 6 Increase 10/300 GL columns | Sigma-Aldrich | CAT# GE29-0915-96 |