Fig. 6.
Effects of TNF-α and suppression of K+ channel activity on NFκB activation. (A) Time-dependent activation of NFκB in response to TNF-α stimulation. HCE cells were simulated with 20 ng/ml TNF-α and then lysed at indicated time points. (B) Dose-dependent effect of suppressing K+ channel activity on TNF-α-induced NFκB activation. Nuclear extracts were displayed in EMSA as described in method. Unlabeled NFκB consensus oligonucleotides (specific competitor), unlabeled SP1 oligonucleotides (nonspecific competitor), anti-NFκB (p65) antibody (for supershift lane), and various concentrations of 4-AP were added to reaction, respectively. (C) Inhibition of TNF-α induced NFκB transnuclear activity by suppressing K+ channel activity. HCE cells were treated with 1 mM 4-AP for 30 min prior to the stimulation of 20 ng/ml TNF-α for 15 min. The cell nuclear and cytosolic extracts were displayed in PAGE.