FIG. 6.

Stable expression of Bcl-2 and Bcl-xL proteins in NT2.N cell lines. Cell lysates prepared from fully differentiated NT2.N neuronal cell lines were subjected to SDS-PAGE and Western blot analysis for Bcl-2, Bcl-xL, Bax, and actin expression. Stable cell lines were created by transfection with either Bcl-2 or Bcl-xL expression plasmids as described in Materials and Methods. NT2.N/neo cells were stably transfected with the plasmid pSV2neo. Equal amounts (25 μg) of protein were loaded into each lane in two separate gels and run in parallel. Membranes were cut to allow separate incubation with Bcl-2, Bcl-xL, Bax-α, and actin MAbs, and the resultant films were analyzed by densitometry to estimate relative protein amounts. (A) Bcl-2 expression. (B) Bcl-xL expression. (C) Bcl-2/Bax-α expression ratio. (D) Bcl-xL/Bax-α expression ratio. Representative results from one of two independent experiments are shown.