Figure 2.

BH3-family transcription and translation is conserved regardless of damage modality. (a) Indicated cell lines were exposed to gemcitabine (10 μM), Cr(VI) (30 μM) or γ-IR (10 Gy) for 48 h. Immunoblotting was carried out for Bcl-2, Bcl-xL, Bid, Bax, PUMA, p53 and β-actin. (b) Cells were treated as indicated and RNA was extracted using Trizol (Invitrogen). cDNA was generated (superscript II, Invitrogen) and qRT-PCR (Sybr Green, Stratagene) was carried out for Bcl-2, Bcl-xL, Bid, Bax, PUMA and GAPDH. Expression was normalized to GAPDH and gene expression (fold change) was calculated using the 2^−ΔΔCT formula. Each assay was conducted in quadruplicate, N=3