FIGURE 7.

The inhibition of MOR mRNA and PARP-1 binding to T⁻¹⁷² probe by the treatment of PARP-1 inhibitor in SH-SY5Y cells. A, total RNA in SH-SY5Y was extracted 24 h after the addition of DMSO, BZD, or BZA or was extracted 72 h after transfection of shRNA expression vector. Then MOR mRNA expression was investigated by RT-PCR. Lane 1, DMSO as a vehicle control (CON); lanes 2–4, BZD (1, 5, and 10 mm); lane 6, BZA as a negative control of BZD; lane 7, control shRNA; lane 8, shPARP-1 (SH-1); lane 9, shPARP-1 (SH-2). PCR cycles of each gene were as follows: MOR, 26 cycles; PARP-1, 25 cycles; DRD2, 28 cycles; glyceraldehyde-3-phosphate dehydrogenase, 20 cycles. B, the real time RT-PCR indicated MOR mRNA expression level normalized to L19 mRNA expression level. The error bars indicate S.D. derived from three independent experiments. The asterisks indicate significantly different values (**, p < 0.01; ***, p < 0.001). C, PARP-1 protein expression detected by WB. SP1 protein expression was used as loading control. D, BZD inhibited PARP-1 binding to the T⁻¹⁷² probe. Nuclear protein in BZD-treated SH-SY5T cells was extracted 24 h after the addition of DMSO (vehicle control) or various concentrations of BZD. Lane 1, nuclear protein treated with DMSO as a vehicle control; lanes 2–4, nuclear protein treated with BZD (1, 5, and 10 mm). These experiments were performed three times or more. I.B., immunoblot.