Figure 4
From: Differential regulation of NF-κB activation and function by topoisomerase II inhibitors
Oxygen free radical generation is not required for NF-κB dependent repression of transcription. (A) U-2 OS cells were transfected with 2 μg of 3 × κB ConA luc and 36 hours after transfection cells were unstimulated (control) or stimulated for 8 hours with 1 μM daunorubicin or 40 J/m2 UV-C, with, or without, 1 hour pre-stimulation with 20 μM EGCG, as indicated. (B) U-2 OS cells were unstimulated (control) or stimulated for 6 hours with 1 μM daunorubicin or 40 J/m2 UV-C, with, or without, 1 hour pre-stimulation with 20 μM EGCG, as indicated. Total RNA was prepared and semi-quantitative PCR analysis with primers specific to human Bcl-xL and GAPDH. (C) Mitoxantrone inhibits Bcl-xL and XIAP mRNA levels. U-2 OS cells were stimulated for 6 hours with 4 μM ICRF-193, 2.5 μM mitoxantrone and 1 μM doxorubicin. Total RNA was prepared and semi-quantitative PCR analysis performed using primers to human Bcl-xL, XIAP and GAPDH control. (D) Quantitative RT-PCR analysis. The effects of danuorubicin, doxorubicin and mitoxantrone on Bcl-xL and XIAP mRNA levels was confirmed by quantitative real-time PCR analysis.