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. Author manuscript; available in PMC: 2008 May 25.
Published in final edited form as: Virology. 2007 Jan 26;362(1):99–108. doi: 10.1016/j.virol.2006.12.020

Figure 3. CD40L is silenced by methylation, but not deacetylation, in Tax-expressing HTLV-I-infected T cell lines.

Figure 3

(A) Real time PCR analysis of CD40L mRNA expression in C8166, MT-2, SLB-1, HUT-102 HTLV-I-infected T cell lines and Jurkat cells non-treated and activated by PMA (10ng/ml)/ionomycin (2μM) treatment. Bars represent relative CD40L mRNA levels normalized to 18S rRNA in the same real time PCR reaction. (B) RT-PCR analysis of CD40L, Tax, and GAPDH expression in C8166 cells treated with or without Trichostatin A (100 nM) for 72 hours and PMA and/or ionomycin for the last 6 hours. (C) Cell surface expression of CD40L protein in C8166 and MT-2 HTLV-I-infected T cell lines with or without treatment with PMA and ionomycin for 6 hours and/or Trichostatin A (500 nM) for 24 hours. Cells were stained with PE conjugated anti-CD40L antibody. Control cells were stained with irrelevant antibody of the same isotype as the anti-CD40L antibody. PE expression was analyzed on 10,000 gated cells using FACScan flow cytometer. (D) RT-PCR analysis of CD40L, Tax, and GAPDH expression in C8166 cells treated with or without 5-aza-2'-deoxycytidine (1 μM) for 72 hours and PMA and/or ionomycin for the last 6 hours. (E) RT-PCR analysis of CD40L, Tax, CD69 and GAPDH in TL-OM1 cells infected with either pCLXSN or pCLXSN-Tax. Cells were treated overnight with either PMA/ionomycin alone, TSA (500 nM) alone or in combination with PMA/ionomycin, 5-aza-dc (1 μm) for 3 days either alone or together with PMA/ionomycin overnight.