Fig. 5
From: A novel positive feedback-loop between the HTLV-1 oncoprotein Tax and NF-κB activity in T-cells
Tax protein is diminished predominantly over Tax transcript levels upon repression of NF-κB activity. a–c Jurkat T-cells were transfected with the Tax expression plasmid pLcXL (LTR-Tax; 20 µg) alone (mock) or together with a dominant negative inhibitor of κB pIκBα-DN (IκBα-DN; 5 µg), replenished to 100 µg DNA with the empty vector pcDNA. At 4 h after transfection, transfected cells were treated, where indicated, for 44 h with 2.5 µM of the IKK2 inhibitor 2-Amino-6-(2-(cyclopropylmethoxy)-6-hydroxyphenyl)-4-(4-piperidinyl)-3-pyridinecarbonitrile (ACHP) or the solvent control DMSO. a Western Blot shows Tax, NF-κB2 p100/p52 and GAPDH. Numbers indicate a representative densitometric analysis of Tax protein expression levels normalized on GAPDH and Tax expression w/o IκBα-DN or the solvent control DMSO. The blot was cut due to technical reasons. b Densitometric analysis of Tax protein, normalized on GAPDH and Tax w/o IκBα-DN or the solvent control DMSO, of three independent experiments was performed. Values ± SD are depicted and were compared using student’s t-test (**p < 0.01). c Tax transcript levels were measured by quantitative PCR (qPCR). Mean relative copy numbers (rcn), normalized on β-actin (ACTB) and Tax w/o IκBα-DN or the solvent control DMSO, of four independent experiments ± SE are shown and were compared using student’s t-test (n.s., not significant). d–f Tax-transformed Tesi cells were treated with 1 µM, 2.5 µM or 5 µM of the IKK2 inhibitor ACHP or the solvent control DMSO. d Western Blot shows Tax, NF-κB2 p100/p52 and GAPDH. Numbers indicate a representative densitometric analysis of Tax protein expression levels normalized on GAPDH and Tax expression in DMSO treated cells. e Densitometric analysis of Tax protein, normalized on GAPDH and Tax in DMSO treated cells, of three independent experiments was performed. Values ± SD are depicted and were compared using student’s t-test (**p < 0.01). f Tax transcript levels were measured by quantitative PCR (qPCR). Mean relative copy numbers (rcn), normalized on β-actin (ACTB) and values derived from DMSO treated cells, of three independent experiments ± SD are shown and were compared using student’s t-test (**p < 0.01)