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. 2013 Apr 16;288(22):15712–15724. doi: 10.1074/jbc.M113.454843

FIGURE 4.

FIGURE 4.

GRK5 is necessary for the CP55940 and GP1a-induced up-regulation of 5-HT2A receptors in CLU213 cells. Cells were transfected with control or GRK5 shRNA lentiviral particles as described under “Experimental Procedures.” A, qRT-PCR to show reduced GRK5 mRNA levels in cells treated with GRK5 shRNA lentiviral particles. B, Western blot to show reduced GRK5 protein levels in cells treated with GRK5 shRNA lentiviral particles. A and B, **, p < 0.01, significant effect of GRK5 shRNA compared with control shRNA-treated cells. C and D, cells stably transfected with control or GRK5 shRNA lentiviral particles were treated with either vehicle, 1 nm CP55940, or 1 nm GP1a for 72 h. C, qRT-PCR to show the effect of GRK5 knockdown on CP55940-induced increases on 5-HT2A receptor mRNA levels. **, p < 0.01, significant effect of CP55940 treatment on 5-HT2A receptor mRNA levels in control shRNA lentivirus-transfected cells compared with vehicle-treated controls. ##, p < 0.01, significant effect of GRK5 shRNA lentivirus transfection on the CP55940-induced up-regulation of 5-HT2A receptors. **, p < 0.01, significant effect of CP55940 treatment in GRK5 shRNA-transfected cells compared with vehicle-treated GRK5 shRNA-transfected cells. D, qRT-PCR to show the effect of GRK5 knockdown on GP1a-induced increases in 5-HT2A receptor mRNA levels. **, p < 0.01, significant effect of GP1a treatment on 5-HT2A receptor mRNA levels in control shRNA lentivirus-transfected cells compared with vehicle-treated controls. ##, p < 0.01, significant effect of GRK5 shRNA lentivirus transfection on the GP1a-induced up-regulation of 5-HT2A receptors. **, p < 0.01, significant effect of GP1a treatment in GRK5 shRNA-transfected cells compared with vehicle-treated GRK5 shRNA-transfected cells. The data represent mean ± S.E. (error bars) (n = 3).