Cell Culture and Viral Propagation

U2OS cells (American Type culture collection (ATCC), Manassas, VA, USA) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Corning Cellgro, Manassas, VA, USA) supplemented with 10% Nu serum (Corning Cellgro), non-essential amino acids (Corning Cellgro), HEPES, sodium pyruvate (Multicell Wisent Inc., St-Bruno, Québec, Canada) and plasmocin 5 μg/ml (InvivoGen, San Diego, CA, USA). Molt-3 (ATCC, CRL-1552), HSB-2 (ATCC, CCL-120.1), Sup-T1 (ATCC, CRL-1942), all human T lymphoblastic cell lines, were cultured in RPMI-1640 (Corning Cellgro) supplemented with 10% fetal bovine serum (FBS), HEPES and plasmocin 5 μg/ml. J-Jhan (RRID:CVCL_1H08) cells infected with wild type (WT) HHV-6A-BAC, HHV-6A mutants lacking telomeric repeats (TMR) termed ΔTMR and ΔimpTMR [6 (link)] or HHV-6A BAC WT#2 (containing a red fluorescent protein downstream of the U11 gene) were cultured in RPMI-1640 supplemented with 10% FBS. HHV-6B (Z29 strain) and HHV-6A (U1102 strain) were propagated and titered on Molt-3 and HSB-2 cells respectively, as previously described [38 (link)].

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Gilbert-Girard S., Gravel A., Collin V., Wight D.J., Kaufer B.B., Lazzerini-Denchi E, & Flamand L. (2020). Role for the shelterin protein TRF2 in human herpesvirus 6A/B chromosomal integration. PLoS Pathogens, 16(4), e1008496.

Publication 2020

U2OS cells DMEM Nu serum non-essential amino acids HEPES plasmocin Molt-3 HSB-2 Sup-T1 RPMI-1640 fetal bovine serum

Cell lines Cells Cultured medium Eagle Essential amino acids Fetal bovine serum Hepes Hhv 2 Hhv 6a Hhv 6b Human Molt Plasmocin Protein gene Pyruvate Serum Sodium Strain Telomeric

Corresponding Organization : Université Laval

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Variable analysis

independent variables

Cell lines used (U2OS, Molt-3, HSB-2, Sup-T1, J-Jhan cells)
HHV-6A and HHV-6B virus strains (WT, ΔTMR, ΔimpTMR, HHV-6A BAC WT#2)

dependent variables

Cell growth and proliferation

control variables

Cell culture media (DMEM, RPMI-1640)
Supplements (10% Nu serum, 10% FBS, non-essential amino acids, HEPES, sodium pyruvate, plasmocin 5 μg/ml)

positive controls

J-Jhan cells infected with wild type (WT) HHV-6A-BAC

negative controls

J-Jhan cells infected with HHV-6A mutants lacking telomeric repeats (ΔTMR and ΔimpTMR)

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