Summary of Study ST002413

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001553. The data can be accessed directly via it's Project DOI: 10.21228/M8ZB01 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST002413
Study Title	IFN-inducible phospholipid levels govern endosomal antiviral immunity
Study Summary	Untargeted lipidomics of 3 cell lines at baseline with focus on phospholipids to understand the role of these lipids in endosomal antiviral immunity
Institute	University of Colorado Denver
Laboratory	Angelo D'Alessandro
Last Name	Haines
First Name	Julie
Address	12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Email	julie.haines@cuanschutz.edu
Phone	3037243339
Submit Date	2022-12-16
Num Groups	3
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2023-01-04
Release Version	1

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Treatment:

Treatment ID:	TR002514
Treatment Summary:	Transduction All cells were transduced at the indicated multiplicity of infection (MOI) as calculated by titration of vector stocks on 293T cells and expressed as transducing units (TU)/293T cell. Cell lines were transduced with LV for 16 hours, washed and kept in culture until reading transduction efficiency by flow cytometry. Human CB-derived HSPC were cultured in serum-free StemSpan medium (StemCell Technologies) supplemented with penicillin (100 IU/ml), streptomycin (100 µg/ml), 100 ng/ml recombinant human stem cell factor (rhSCF), 20 ng/ml recombinant human thrombopoietin (rhTPO), 100 ng/ml recombinant human Flt3 ligand (rhFlt3), and 20 ng/ml recombinant human IL6 (rhIL6) (all from Peprotech) 16 to 24 hours prior to transduction. HSPC were then transduced at a concentration of 1×106 cells per milliliter with VSV-gp-pseudotyped SINLV for 16 hours at the indicated multiplicity of infection (MOI) in the same medium. G-CSF mobilized peripheral blood CD34+ cells were maintained in culture in CellGro medium (Cell Genix) containing a cocktail of cytokines: 60 ng/ml IL-3, 100 ng/ml TPO, 300 ng/ml SCF, and 300 ng/ml FLT-3L (all from Cell Peprotech). Transductions were performed with the indicated dose of vectors for 16 hours in the same cytokine-containing medium. For single hit reporter LV transductions cells were washed and maintained in serum-free medium supplemented with cytokines as above until the reading of the percentage of positive cells by FACS. To perform OE, KD and KO experiments cells were transduced with the respective LV and then challenged with a second transduction with or without drugs. Except for primary cells, KD and KO cells were selected with Puromycin before the second hit of transduction.

Treatment ID:

TR002514

Treatment Summary:

Transduction All cells were transduced at the indicated multiplicity of infection (MOI) as calculated by titration of vector stocks on 293T cells and expressed as transducing units (TU)/293T cell. Cell lines were transduced with LV for 16 hours, washed and kept in culture until reading transduction efficiency by flow cytometry. Human CB-derived HSPC were cultured in serum-free StemSpan medium (StemCell Technologies) supplemented with penicillin (100 IU/ml), streptomycin (100 µg/ml), 100 ng/ml recombinant human stem cell factor (rhSCF), 20 ng/ml recombinant human thrombopoietin (rhTPO), 100 ng/ml recombinant human Flt3 ligand (rhFlt3), and 20 ng/ml recombinant human IL6 (rhIL6) (all from Peprotech) 16 to 24 hours prior to transduction. HSPC were then transduced at a concentration of 1×106 cells per milliliter with VSV-gp-pseudotyped SINLV for 16 hours at the indicated multiplicity of infection (MOI) in the same medium. G-CSF mobilized peripheral blood CD34+ cells were maintained in culture in CellGro medium (Cell Genix) containing a cocktail of cytokines: 60 ng/ml IL-3, 100 ng/ml TPO, 300 ng/ml SCF, and 300 ng/ml FLT-3L (all from Cell Peprotech). Transductions were performed with the indicated dose of vectors for 16 hours in the same cytokine-containing medium. For single hit reporter LV transductions cells were washed and maintained in serum-free medium supplemented with cytokines as above until the reading of the percentage of positive cells by FACS. To perform OE, KD and KO experiments cells were transduced with the respective LV and then challenged with a second transduction with or without drugs. Except for primary cells, KD and KO cells were selected with Puromycin before the second hit of transduction.