Fig. 3
From: A novel positive feedback-loop between the HTLV-1 oncoprotein Tax and NF-κB activity in T-cells
Tax protein levels are elevated independent of the promoter. a Schematic representation of the Tax expression constructs and their promoters used in (b–d). b–d Jurkat T-cells were transiently transfected with the Tax expression constructs pc-Tax, pCAG-FLAG-Tax (pCAG-Tax), pSG5-Tax, pEF-Tax1 or pLcXL (LTR-Tax; 30 µg each) and the IKK2 constitutive active mutant pcFLAG-IKK2-EE (FLAG-IKK2-EE) or the empty vector (pcDNA; 40 µg each). All samples were replenished to 100 µg with empty vector pcDNA. b Western Blot shows Tax, NF-κB2 p100/p52, FLAG and GAPDH as controls. Numbers depict a representative densitometric analysis of Tax protein expression levels normalized on GAPDH and Tax expression w/o IKK2-EE. c Tax transcript levels were measured by quantitative PCR (qPCR). Mean relative copy numbers (rcn), normalized on β-actin (ACTB) and on Tax w/o IKK2-EE, of four independent experiments ± SE are shown and were compared using student’s t-test (*p < 0.05). The inset shows an enlargement of Tax transcript values derived from pCAG-Tax and pSG5-Tax transfected Jurkat T-cells. d Correlation between Tax protein values, derived from four independent experiments as depicted in (b), and their respective transcript levels as depicted in (c) was calculated using Pearson’s correlation (R2 = 0.3982; p > 0.05)