Tumor cells (2 × 106) were prepared for the chromatin immunoprecipitation (ChIP) assay with the ChIP assay kit (Cell Signaling Technology). The CREB-bound DNA of the GLUT3 gene was immunoprecipitated with CREB antibodies. The DNA was then used for polymerase chain reaction (PCR) using primers designed for the amplification of the GLUT3 promoter. The PCR product was analyzed and quantified as described before [24 (link)]. The primers used were as follows:
(1) forward: 5′-TATTTTCTT CTCCTGCTTAGCT-3′ and
reverse: 5′-AGTCATT TATAGT GTTTCCCTTC-3′ and
(2) forward: 5′-CCCAGGGTGGA GAGAGTGGAAG-3′ and
reverse 5′-TTATAATCTCCGCAA AGGGTGGAG-3′ and
(3) forward: 5′-GTCATATCCC AGCGAGACCC AG-3′ and
reverse: 5′-C GCTGTAATCTAA TTCAAGTCTTCAAG-3′.
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