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Fig. 2 | Retrovirology

Fig. 2

From: A novel positive feedback-loop between the HTLV-1 oncoprotein Tax and NF-κB activity in T-cells

Fig. 2

Protein expression of the NF-κB deficient Tax mutants M22 and K1-10R can be rescued by co-expression of IKK2-EE in various T-cell lines. a, b, e, f Jurkat T-cells or c, d CCRF-CEM and Molt-4 T-cells were transfected with the Tax wildtype expression plasmid (ad) pEF-Tax1 (Tax, 30 µg) or (e, f) pSG5-Tax (Tax, 5 µg) and the NF-κB-deficient Tax mutant (a, b) pEF-M22 (M22, 30 µg) or (e, f) pSG5-Tax-K1-10R (K1-10R, 15 µg) together with the IKK2 constitutive active mutant pcFLAG-IKK2-EE (FLAG-IKK2-EE, 40 µg) or the empty vector (pcDNA). All samples were replenished to 100 µg with the respective empty vector DNA (pcDNA, pSG5). a, c, e Immunoblot shows Tax, NF-κB2 p100/p52, FLAG or IKK2 and ACTB or GAPDH as controls. Numbers depict a representative densitometric analysis of Tax protein expression levels normalized on ACTB or GAPDH and Tax expression w/o IKK2-EE. b, d, f Densitometric analysis of Tax protein expression relative to ACTB or GAPDH, normalized on Tax expression w/o IKK2-EE, of at least three independent experiments was performed (protein, black bars). Values ± SD are depicted. Corresponding Tax transcript levels were measured by quantitative PCR (qPCR; transcripts, grey bars). Mean relative copy numbers (rcn), normalized on β-actin (ACTB) and on Tax w/o IKK2-EE, of three independent experiments ± SD were compared using student’s t-test (*p < 0.05; n.s. not significant). The inset in (b) shows an enlargement of Tax protein and Tax transcript levels derived from Tax wildtype transfected Jurkat T-cells

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