HTLV-1-Infected T-Cell Lines and Assays

HTLV-1-infected T-cell lines (ED-40515(−), MT-2, Hut-102, and ATL-43T) and non–HTLV-1–infected T-cell line (Jurkat T cell) were cultured in RPMI1640 (Nacalai Tesque) containing 10% FBS and 1% PSG (Invitrogen). Jurkat T cells stably expressing a spliced form of HBZ called Jurkat-HBZ were maintained as described previously^{44 (link)}. HEK293T cells were maintained with DMEM (Nacalai Tesque) containing 10% FBS and 1% PSG. The Jurkat Tet/On-Tax cells^{45 (link)} kindly gifted from Dr. W.C. Greene were maintained in RPMI1640 supplemented with 10% tetracycline-free FBS (Clontech) and treated with 1 µg/ml doxycycline for 36 hours for inducible Tax expression. Human-cell-killing assay was performed as described previously^{9 (link)}.

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Takiuchi Y., Kobayashi M., Tada K., Iwai F., Sakurada M., Hirabayashi S., Nagata K., Shirakawa K., Shindo K., Yasunaga J.I., Murakawa Y., Rajapakse V., Pommier Y., Matsuoka M, & Takaori-Kondo A. (2017). HTLV-1 bZIP factor suppresses TDP1 expression through inhibition of NRF-1 in adult T-cell leukemia. Scientific Reports, 7, 12849.

Publication 2017

RPMI1640 PSG DMEM tetracycline-free FBS

Assay Cell Cell line Doxycycline Htlv Human Jurkat cells Tetracycline

Corresponding Organization :

Other organizations : Kyoto University, RIKEN Center for Integrative Medical Sciences, National Cancer Institute, Center for Cancer Research, National Institutes of Health, Kumamoto University

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Variable analysis

independent variables

Presence of HTLV-1 infection in T-cell lines (ED-40515(-), MT-2, Hut-102, and ATL-43T)
Presence of spliced form of HBZ expression in Jurkat T cells
Induction of Tax expression in Jurkat Tet/On-Tax cells with doxycycline

dependent variables

Outcome of human-cell-killing assay

control variables

Culture medium (RPMI1640 containing 10% FBS and 1% PSG) for HTLV-1-infected T-cell lines and non-HTLV-1-infected Jurkat T cells
Culture medium (DMEM containing 10% FBS and 1% PSG) for HEK293T cells
Culture medium (RPMI1640 supplemented with 10% tetracycline-free FBS) for Jurkat Tet/On-Tax cells

controls

Non-HTLV-1-infected Jurkat T cell line as a negative control
Jurkat T cells stably expressing a spliced form of HBZ as a positive control

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