Journal: Immunity

Article Title: m 6 A Modification Prevents Formation of Endogenous Double-Stranded RNAs and Deleterious Innate Immune Responses during Hematopoietic Development

doi: 10.1016/j.immuni.2020.05.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cat# 10010200; RRID: AB_2651015 Cy3 Streptavidin VECTOR LABORATORIES Cat# SA-1300 Cy5 Streptavidin VECTOR LABORATORIES Cat# SA-1500 Chemicals, Peptides, and Recombinant Proteins MethoCult™ GF M3434 STEMCELL Technologies Cat# 03434 Phusion High-Fidelity PCR Master Mix with HF Buffer New England Biolabs Cat# M0531L RBC Lysis Buffer, 10X Santa Cruze Biotechnology Cat# sc-296258 Lipofectamine™ 2000 Transfection Reagent ThermoFisher SCIENTIFIC Cat# 11668019 Lipofectamine™ RNAiMAX Transfection Reagent ThermoFisher SCIENTIFIC Cat# 13778100 Mouse Thrombopoietin (mTPO) Gemini Bio-product Cat# 300-351P Mouse Stem Cell Factor (mSCF) Gemini Bio-product Cat# 300-348P Mouse Flt-3 Ligand (mFlt3L) Gemini Bio-product Cat# 300-306P Mouse Interleukin-3 (mIL3) Gemini Bio-product Cat# 300-324P Benchmark Fetal Bovine Serum (FBS) Gemini Bio-product Cat# 100-106 DMEM, high glucose ThermoFisher SCIENTIFIC Cat# 11965092 Protein A–micrococcal nuclease (pA-MN) fusion protein Henikoff lab Skene and Henikoff, 2018 Spermidine trihydrochloride Sigma-Aldrich Cat# S2501 Con A-coated BioMag®Plus microparticles Polysciences, Inc Cat# 86057-3 iQ™ SYBR® Green Supermix BIO-RAD Cat# 1708880 iScript cDNA Synthesis Kit BIO-RAD Cat# 1708890 cOmplete™, Mini, EDTA-free Protease Inhibitor Cocktail Sigma-Aldrich Cat# 11836170001 SUPERase• In™ RNase Inhibitor ThermoFisher SCIENTIFIC Cat# AM2696 SPRIselect beads Beckman Coulter Cat# {"type":"entrez-nucleotide","attrs":{"text":"B23317","term_id":"2508948","term_text":"B23317"}} B23317 RNaseA, Dnase and protease-free ThermoFisher SCIENTIFIC Cat# EN0531 Maxima H Minus Reverse Transcriptase ThermoFisher SCIENTIFIC Cat# EP0752 Exonuclease I New England Biolabs Cat# M0293S Ampure XP beads Beckman Coulter Cat# A63880 Fluorinert™ FC-40 Sigma-Aldrich Cat# F9755 Critical Commercial Assays EpiQuik m 6 A RNA Methylation Quantification Kit (Colorimetric) Epigentek Cat# P-9005-96 FITC BrdU Flow Kits BD Pharmingen™ Cat# 559619 CellTrace CFSE Cell Proliferation Kit, for flow Thermo Fisher Cat# {"type":"entrez-nucleotide","attrs":{"text":"C34554","term_id":"2370695","term_text":"C34554"}} C34554 FITC Annexin V Apoptosis Detection Kit I BD Pharmingen™ Cat# 556547 RNeasy Mini Kit QIAGEN Cat# 74106 RNeasy Plus Micro Kit QIAGEN Cat# 74034 Magnetic mRNA Isolation Kit New England Biolabs Cat# S1550S MinElute PCR Purification Kit QIAGEN Cat# 28004 NEBNext Multiplex Oligos for Illumina New England Biolabs Cat# E7500S NEBNext Ultra II DNA Library Prep Kit New England Biolabs Cat# E7645S Qubit dsDNA HS Assay Kit Life Technologies Cat# {"type":"entrez-protein","attrs":{"text":"Q32851","term_id":"75280859","term_text":"Q32851"}} Q32851 Vector M.O.M.

Techniques: Purification, Blocking Assay, Plasmid Preparation, Recombinant, Lysis, Transfection, SYBR Green Assay, Protease Inhibitor, Methylation, Isolation, Multiplex Assay, Immunodetection, Software, Sequencing

Journal: Immunity

Article Title: m 6 A Modification Prevents Formation of Endogenous Double-Stranded RNAs and Deleterious Innate Immune Responses during Hematopoietic Development

doi: 10.1016/j.immuni.2020.05.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cat# 10010200; RRID: AB_2651015 Cy3 Streptavidin VECTOR LABORATORIES Cat# SA-1300 Cy5 Streptavidin VECTOR LABORATORIES Cat# SA-1500 Chemicals, Peptides, and Recombinant Proteins MethoCult™ GF M3434 STEMCELL Technologies Cat# 03434 Phusion High-Fidelity PCR Master Mix with HF Buffer New England Biolabs Cat# M0531L RBC Lysis Buffer, 10X Santa Cruze Biotechnology Cat# sc-296258 Lipofectamine™ 2000 Transfection Reagent ThermoFisher SCIENTIFIC Cat# 11668019 Lipofectamine™ RNAiMAX Transfection Reagent ThermoFisher SCIENTIFIC Cat# 13778100 Mouse Thrombopoietin (mTPO) Gemini Bio-product Cat# 300-351P Mouse Stem Cell Factor (mSCF) Gemini Bio-product Cat# 300-348P Mouse Flt-3 Ligand (mFlt3L) Gemini Bio-product Cat# 300-306P Mouse Interleukin-3 (mIL3) Gemini Bio-product Cat# 300-324P Benchmark Fetal Bovine Serum (FBS) Gemini Bio-product Cat# 100-106 DMEM, high glucose ThermoFisher SCIENTIFIC Cat# 11965092 Protein A–micrococcal nuclease (pA-MN) fusion protein Henikoff lab Skene and Henikoff, 2018 Spermidine trihydrochloride Sigma-Aldrich Cat# S2501 Con A-coated BioMag®Plus microparticles Polysciences, Inc Cat# 86057-3 iQ™ SYBR® Green Supermix BIO-RAD Cat# 1708880 iScript cDNA Synthesis Kit BIO-RAD Cat# 1708890 cOmplete™, Mini, EDTA-free Protease Inhibitor Cocktail Sigma-Aldrich Cat# 11836170001 SUPERase• In™ RNase Inhibitor ThermoFisher SCIENTIFIC Cat# AM2696 SPRIselect beads Beckman Coulter Cat# {"type":"entrez-nucleotide","attrs":{"text":"B23317","term_id":"2508948","term_text":"B23317"}} B23317 RNaseA, Dnase and protease-free ThermoFisher SCIENTIFIC Cat# EN0531 Maxima H Minus Reverse Transcriptase ThermoFisher SCIENTIFIC Cat# EP0752 Exonuclease I New England Biolabs Cat# M0293S Ampure XP beads Beckman Coulter Cat# A63880 Fluorinert™ FC-40 Sigma-Aldrich Cat# F9755 Critical Commercial Assays EpiQuik m 6 A RNA Methylation Quantification Kit (Colorimetric) Epigentek Cat# P-9005-96 FITC BrdU Flow Kits BD Pharmingen™ Cat# 559619 CellTrace CFSE Cell Proliferation Kit, for flow Thermo Fisher Cat# {"type":"entrez-nucleotide","attrs":{"text":"C34554","term_id":"2370695","term_text":"C34554"}} C34554 FITC Annexin V Apoptosis Detection Kit I BD Pharmingen™ Cat# 556547 RNeasy Mini Kit QIAGEN Cat# 74106 RNeasy Plus Micro Kit QIAGEN Cat# 74034 Magnetic mRNA Isolation Kit New England Biolabs Cat# S1550S MinElute PCR Purification Kit QIAGEN Cat# 28004 NEBNext Multiplex Oligos for Illumina New England Biolabs Cat# E7500S NEBNext Ultra II DNA Library Prep Kit New England Biolabs Cat# E7645S Qubit dsDNA HS Assay Kit Life Technologies Cat# {"type":"entrez-protein","attrs":{"text":"Q32851","term_id":"75280859","term_text":"Q32851"}} Q32851 Vector M.O.M.

Techniques: Purification, Blocking Assay, Plasmid Preparation, Recombinant, Lysis, Transfection, SYBR Green Assay, Protease Inhibitor, Methylation, Isolation, Multiplex Assay, Immunodetection, Software, Sequencing

Journal: Molecular Medicine Reports

Article Title: Complement factor H-related 3 overexpression affects hepatocellular carcinoma proliferation and apoptosis

doi: 10.3892/mmr.2019.10514

Figure Lengend Snippet: Effect of CFHR3 on Huh-7 cell proliferation and apoptosis. (A-C) Cell proliferation was measured by flow cytometric analysis of CFSE. (D) Quantification of the cell proliferation data. (E-G) Apoptosis was detected by flow cytometric analysis of Annexin V-FITC- and PI-stained cells. (H) Quantification of the apoptosis rate. Data are presented as the mean ± standard deviation from three independent experiments. **P<0.01 vs. Con. Con, control; NC, negative control; CFHR3, complement factor H-related 3; CFSE, carboxyfluorescein succinimidyl ester; FITC, fluorescein isothiocyanate; PI, propidium iodide.

Article Snippet: Cells were labeled with CellTrace TM CFSE kit (C34554; Invitrogen; Thermo Fisher Scientific, Inc.) for 10 min at 37°C and then washed twice with phosphate-buffered saline.

Techniques: Staining, Standard Deviation, Control, Negative Control