FIG 8.

Relationship between IFN effects and Pde4dip effects. (A) Pde4dip mRNA levels in N167 cells treated with IFN-α. Cells were treated with 5 μg/ml Gly-9 (Gly-9 +) or vehicle (Gly-9 −) for 3 days and, simultaneously, with 5 U/ml IFN-α for the indicated periods before the harvest. Data are averages and standard deviations for triplicate experiment results (n.s., not significant; **, P < 0.05 versus vehicle control of IFN-α). (B) Temporal profiles of mRNA levels of representative IFN-stimulated genes in N167 cells treated with si-Pde4dip. The mRNA levels of Cxcl10 and Ifit1 were analyzed in cells treated with combinations of 5 μg/ml Gly-9 and 10 nM si-Pde4dip. Gly-9 treatment started half a day before transfection of si-Pde4dip. Cells were harvested at the indicated time points after transfection. The vehicle for Gly-9 and the transfection reagent for si-Pde4dip were used as negative controls. Data are averages and standard deviations for triplicate experimental results (n.s., not significant; *, P < 0.01; **, P < 0.05). (C) Immunoblotting of PrPres in N167 cells treated with combinations of si-Pde4dip and IFN-α. Cells in the presence or absence of 5 μg/ml Gly-9 were treated with 5 U/ml IFN-α for three and a half days and, simultaneously, with 10 nM si-Pde4dip for the last 3 days before harvesting. The vehicles for Gly-9 and IFN-α and the transfection reagent for si-Pde4dip were used as negative controls. Graphic data are averages and standard deviations for triplicate immunoblot signals (n.s., not significant; *, P < 0.01; **, P < 0.05).