Bcl 2

Forty-eight hours after transfection, cells were collected and total RNA was extracted using TRIzol (Invitrogen, Carlsbad, California, USA). For RT-PCR, RNA was reverse transcribed with M-MuLV (Fermentas, Vilnius, Lithuania) to form cDNA. The primer pairs for the PCR amplification of Bax, Bcl-2 and GAPDH (glyceraldehyde 3-phosphate dehydrogenase) were synthesized by Sangon Biotech Co., Ltd (Shanghai, China) and their sequences are given in Table 1.
Table 1.

Primer sequences used in reverse transcription-PCR.

Bax	F1: 5′AAGCTGAGCGAGTGTCTCAAG3′
	R1: 5′CAAAGTAGAAAGGGCGACAAC3′
Bcl-2	F1: 5′TGGGAGAACAGGGTACGATAAC3′
	R1: 5′GAACTCAAAGAAGGCCACAATC3′
GAPDH	F1: 5′GGGAAACTGTGGCGTGAT3′
	R1: 5′GAGTGGGTGTCGCTGTTGA3′

DNA polymerase rTaq (TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, China) was used to amplify Bax, Bcl-2 and GAPDH from the template cDNA. GAPDH was used as an internal control. The PCR products of Bax, Bcl-2 and GAPDH were analysed in a 1.5% agarose gel and scanned by a gel-imaging system (Shanghai Forte Co., Shanghai, China).