RNA from treated or untreated cells was extracted using Trizol (Thermo Fisher Scientific, MA USA) following manufacturer’s instructions. 200 ng of total RNA from each sample were retro-transcribed using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystem, Thermo Fisher Scientific, MA USA). qPCR reactions were performed by means of a 7900 HT Real Time PCR (Applied Biosystem) using gene specific primers for the following selected genes:
BAX: Forward 5′-TTTGCTTCAGGGTTTCATCCA-3′: Reverse 5′- CTCCATGTTACTGTCCAGTTCGT-3′; BCL-2: Forward 5′-GTTCCCTTTCCTTCCATCC-3′; Reverse 5′-TAGCCAGTCCAGAGGTGAG-3′; FAS: Forward 5′-CCCTCCTACCTCTGGTTCTTACG-3′; Reverse 5’TCAGTCACTTGGGCATTAACACTTT-3′; FASL: Forward 5′-CCTGAAAAAAAGGAGCTGAGGAA-3′; Reverse 5′-GGCATGGACCTTGAGTTGGA-3′; GAPDH: Forward 5′-CAAGGCTGTGGGCAAGGT-3′; Reverse 5′-GGAAGGCCATGCCAGTGA-3:
Primers were designed at exon-exon junctions using Primer express 2.0 (Applied Biosystems). Target expression level was performed as previously described [19 (link)] using GAPDH as housekeeping gene. All the experiments were performed in triplicate. Data are expressed as the mean ± SD.
BAX: Forward 5′-TTTGCTTCAGGGTTTCATCCA-3′: Reverse 5′- CTCCATGTTACTGTCCAGTTCGT-3′; BCL-2: Forward 5′-GTTCCCTTTCCTTCCATCC-3′; Reverse 5′-TAGCCAGTCCAGAGGTGAG-3′; FAS: Forward 5′-CCCTCCTACCTCTGGTTCTTACG-3′; Reverse 5’TCAGTCACTTGGGCATTAACACTTT-3′; FASL: Forward 5′-CCTGAAAAAAAGGAGCTGAGGAA-3′; Reverse 5′-GGCATGGACCTTGAGTTGGA-3′; GAPDH: Forward 5′-CAAGGCTGTGGGCAAGGT-3′; Reverse 5′-GGAAGGCCATGCCAGTGA-3:
Primers were designed at exon-exon junctions using Primer express 2.0 (Applied Biosystems). Target expression level was performed as previously described [19 (link)] using GAPDH as housekeeping gene. All the experiments were performed in triplicate. Data are expressed as the mean ± SD.
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