Complete NF-κB inactivation induces rapid accumulation of Bcl-xS in quiescent human blood granulocytes.
(A) Blood granulocytes were isolated from buffy coats by density centrifugation and cultured for 2 hours, 4 hours, and 6 hours in the presence or absence of 1.5 μM mGTX, 1.5 μM GTX, or 48 μM PGA1. Whole-cell extracts were prepared and analyzed by immunoblotting for Bax, Bcl-2, Bfl-1, and Bcl-xL/S expression (B). RNA was prepared from blood granulocytes cultured for 2 hours and 4 hours in the presence or absence of 1.5 μM mGTX or 1.5 μM GTX and analyzed by RT-PCR for expression of bcl-xL/S. To control for quantification, GAPDH was also amplified. Filled columns show the amount of total bcl-x mRNA and the ratio betweenbcl-xS and bcl-xL mRNAs, as determined by densitometry analyses. These results represent at least 3 comparable experiments.
