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. 2011 Jan 5;286(10):7854–7864. doi: 10.1074/jbc.M110.160911

FIGURE 1.

FIGURE 1.

Down-regulation of RGS4 protein by overnight treatment with DAMGO or DPDPE. Left panels, representative Western blots of RGS4 protein expression in SH-SY5Y cells are shown. Right panels, combined data are shown from experiments quantified using a Kodak image station. A, whole cell lysates were prepared from SH-SY5Y cells treated with either DAMGO or DPDPE (10 μm, overnight) and subjected to 12% SDS-PAGE. RGS4 was identified with RGS4-specific antibody as described under “Experimental Procedures.” RGS4 protein expression was decreased in DAMGO (DA; ***, p < 0.001)- and DPDPE (DP; ***, p < 0.001)-treated cells, compared with the control (Ctr) without agonist treatment (n = 9). The RGS4 standard (Std) was lysates of HEK293T cells overexpressing RGS4 protein. β-Actin as loading control was not changed. B, naloxone (NLX, 10 μm, overnight (o/n), n = 3) prevented the DAMGO- and DPDPE-induced down-regulation of RGS4 protein. C, preincubation of SH-SY5Y cells for 6 h with PTX (100 ng/ml) followed by overnight incubation in the absence or presence of DAMGO or DPDPE (10 μm) completely blocked DAMGO- or DPDPE-induced down-regulation of RGS4 protein (n = 3, ***, p < 0.001); treatment with PTX alone did not change the level of RGS4 protein compared with non-treated cells (Ctr).