Recombinant:Article Title: N 7 -Methylguanosine tRNA modification enhances oncogenic mRNA translation and promotes intrahepatic cholangiocarcinoma progression.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-METTL1 Proteintech Cat# 14994-1-AP Mouse monoclonal anti-EGFR Proteintech Cat# 66455-1-IG Rabbit polyclonal anti-GAPDH Proteintech Cat# 10494-1-AP Rabbit monoclonal anti-WDR4 Abcam Cat# ab169526 Rabbit monoclonal anti-CK19 Abcam Cat# ab52625 Rabbit polyclonal anti-Ki67 Abcam Cat# ab15580 Mouse monoclonal anti-CCNA2 CST Cat# 4656S Rabbit monoclonal anti-CDK6 CST Cat# 13331S Rabbit polyclonal anti-CDK8 CST Cat# 4106S Rabbit polyclonal anti-mTOR CST Cat# 2972S Rabbit polyclonal anti-pmTOR CST Cat# 2974S Rabbit monoclonal anti-AKT CST Cat# 4691S Rabbit monoclonal anti-pAKT CST Cat# 4060S Anti-rabbit IgG HRP-linked Antibody CST Cat# 7074S Anti-mouse IgG HRP-linked Antibody CST Cat# 7076S Rabbit polyclonal anti-beta-Actin Abbkine Cat# a01011 Anti-Digoxigenin-AP Fab fragments Roche Cat# 11093274910 Mouse monoclonal 7- methylguanosine (m7G) MBL International Cat# RN017M Mouse monoclonal anti-puromycin Millipore Cat# MABE343 Biological samples Human intrahepatic cholangiocarcinoma tissue The First Affiliated Hospital of Sun Yat-sen University https://lt.medidata.cn Chemicals, peptides, and recombinant proteins RPMI Medium 1640 basic (1X) GIBCO Cat# C11875500BT Fetal bovine serum GIBCO Cat# A3160801 Penicillin-Streptomycin GIBCO Cat# 15140163 Lipofectamine 3000 Invitrogen Cat# L3000015 Polybrene Solarbio Cat# H8761 Puromycin Solarbio Cat# P8230 Trizol reagent Life technologies Cat# 15596018 Cycloheximide MedchemExpress Cat# HY-12320 MOPS BioFroxx Cat# 1173GR100 MgCl2 Aladdin Cat# M113687 NaCl Aladdin Cat# C111549 TritonTM X-100 Sigma Cat# 78787 RNasin Ribonuclease Inhibitors Invitrogen Cat# 10777-019 Heparin Aladdin Cat# D130947 PMSF Roche Cat# 10837091001 Benzamidine Sigma Cat# 12072 Sodium borohydride (NaBH4) Sigma Cat# 452882 Aniline Sigma Cat# 242284 HEPES-KOH Bestbio Cat# BB-19324 (Continued on next page) e1 Molecular Cell 81, 3339–3355.e1–e8, August 19, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER KCl Aladdin Cat# P112134 Dithiothreitol Aladdin Cat# D104859 Critical commercial assays mirVana miRNA isolation Kit invitrogen Cat# AM1561 Oligo Clean & Concentrator ZYMO Research Cat# D4061 Cell Cycle Assay Kit PI/Rnase staining Dojindo laboratories Cat# C543 RealTM Envision TM Detection System Peroxidase/DAB+,Rabbit/Mouse Dako Cat# K5007 Cell Counting Kit-8 Dojindo laboratories Cat# CK04 Multiplex Small RNA Library Prep set for Illumina Kit New England Biolabs Cat# E7580S PrimeScript RT reagent Kit Takara Cat# RR036A-1 TB GreenTM Premix Ex TaqTM II Takara Cat# RR820A Mouse Tail Direct PCR Kit Foregene Cat# TP-0134 BrdU Cell Proliferation Kit for Flow Cytometry Manual BioLegend Cat# 370704 Dual-Luciferase Reporter Assay System Promega Cat# E1910 Deposited data RNC-seq Data This paper GEO accession number: 161319 m7G-TRAC-Seq Data This paper GEO accession number: 161319 Ribo-seq Data This paper GEO accession number: 161319 Experimental models: Cell lines HuCCT1 Japan Health Sciences Foundation Resources Bank Cat# JCRB0425 RBE Chinese Type Culture Collection Cat# 31800022 Experimental models: Organisms/strains Mouse: C57BL/6Gpt GemPharmatech GemPharmatech: N000013 Mouse: NOD/ShiLtJGptPrkdcem26Cd52Il2rgem26Cd22/Gpt GemPharmatech GemPharmatech: T001475 Mouse: C57BL/6-Mettl1fl/fl Biocytogen EGE-ZLF-009 Mouse: B6.Cg-Speer6-ps1Tg(Alb-cre)21Mgn/J Jackson Laboratory Cat# 003574 Oligonucleotides siRNA targeting 30UTR sequence: METTL1 #1: CCACTGGTGTTCATGACTA Ribobio N/A siRNA targeting 30UTR sequence: METTL1 #2: AGCTGGAATGAGGAATGTA Ribobio N/A siRNA targeting sequence: METTL1 #1: GATGACCCAAAGGATAAGAAA Ribobio N/A siRNA targeting sequence: METTL1 #2: GGATGTGCACTCATTTCGA Ribobio N/A Probe: tRNA-LysCTT:CGACCCTGAGATTAAGAGTC Sangon Biotech N/A Probe: tRNA-LysTTT:TAAAAGTCTGATGCTCTACC Sangon Biotech N/A Probe: tRNAGlyCCC:GTCTCCCACGTGGGAGGCGA Sangon Biotech N/A Probe: U6 snoRNA:TGGAACGCTTCACGAATTTG Sangon Biotech N/A Primers for qPCR analysis see Table S5 TIANYI HUIYUAN N/A (Continued on next page) Molecular Cell 81, 3339–3355.e1–e8, August 19, 2021 e2
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