Journal: Journal of Hepatocellular Carcinoma

Article Title: Re-Recognizing the Cellular Origin of the Primary Epithelial Tumors of the Liver

doi: 10.2147/JHC.S334935

Figure Lengend Snippet: Monoclonal Antibodies Used in This Study

Article Snippet: Mouse anti-human LFABP , 1:100 , L2B10 , OriGene Tech. , Rockville, MA, USA , AM26273PU-N.

Techniques: Bioprocessing

Journal: bioRxiv

Article Title: In-vivo targeted tagging of RNA isolates cell specific transcriptional responses to environmental stimuli and identifies liver-to-adipose RNA transfer

doi: 10.1101/670398

Figure Lengend Snippet: A) HD5EU small molecule and metabolite structure relative to 5EU. B) W.B. depicting tissue expression pattern of CYP3A4 in humanize CYP3A4 cluster deleted mice. HSP90 serves as loading control. GAS = Gastrocnemius muscle; CM = Cardiac Muscle; BAT = Brown adipose tissue; ScA = Subcutaneous white adipose tissue; VsA = Visceral white adipose tissue; S.I. = Small intestine; L.I. = Large intestine. C) qRT-PCR validating liver and kidney specific expression of CYP3A4. Error-bars for standard-deviation of 3 biological repeats. D) Administration of the HD5EU small molecule to cells expressing CYP3A4 allows metabolism to 5EU and labelling of total RNA. Labeled transcripts are then secreted to the extracellular matrix and can be identified in bio-fluids and recipient cells.

Article Snippet: In addition to the HD5EU small molecule we took advantage of the published humanized liver specific CYP3A4 mouse line FVB/129P2- Cyp3a13 tm1Ahs Del( 5Cyp3a57-Cyp3a59 ) 1Ahs Tg( APOE -CYP3A4) A1Ahs obtained from Taconic [ , ].

Techniques: Expressing, Quantitative RT-PCR, Standard Deviation, Labeling

Journal: bioRxiv

Article Title: In-vivo targeted tagging of RNA isolates cell specific transcriptional responses to environmental stimuli and identifies liver-to-adipose RNA transfer

doi: 10.1101/670398

Figure Lengend Snippet: A) Extracted ion chromatograms from A, C, G, U and 5EU and gradient slope. B) Close up of extracted ion chromatograms of 5EU [M+H] + and 5EU in-source fragment [M-132+H] + . C) Tandem MS spectra at 20 eV of standard and sample indicating main fragment [M-132+H] + . D) Further fragments at 40eV used for identification of 5EU in RNA samples. A-D upper panels depict standard, lower panels depict liver long RNA from HD5EU treated mice. E) Relative quantification of 5EU in primary hepatocytes treated for 8h with the indicated compounds. Azamulin treatment inhibits CYP3A4 mediated HD5EU metabolism to 5EUMP and its subsequent incorporation into RNA. F) Bioanalyzer image depicting isolation of liver long and short RNA. G) Relative quantification of 5EU in liver derived long / small RNA following indicated time after 5EU / HD5EU administration to mice. Error bars indicated standard deviation calculated for 3 biological replicates.

Article Snippet: In addition to the HD5EU small molecule we took advantage of the published humanized liver specific CYP3A4 mouse line FVB/129P2- Cyp3a13 tm1Ahs Del( 5Cyp3a57-Cyp3a59 ) 1Ahs Tg( APOE -CYP3A4) A1Ahs obtained from Taconic [ , ].

Techniques: Isolation, Derivative Assay, Standard Deviation