Figure 6.

PD174265 reverses EGF or TGF driven increase in PS-ASO activity. (A) The levels of Drosha and Malat1 RNAs were quantified by qRT-PCR analysis of A431 cells treated with EGF or EGF and PD174265. Data are relative to untreated cells. The error bars represent standard deviations from 3 independent experiments. Activity curves were fitted from data plotted in panels based on a non-liner regression model and IC₅₀ was calculated based on a non-linear regression model. P (in red) <0.01, EGF or TGF treatment versus control; P (in red) <0.01, EGF treatment versus control; P values were computed by one-way ANOVA as the concentrations of PS-ASOs were set as random effect. (B) Western analyses of proteins in A431 cells pre-treated with or without PD174265 followed by the treatment of EGF. The blot was probed sequentially with different antibodies for phosphorylated EGFR (P-EGFR), total EGFR (T-EGFR), phosphorylated ERK (P-ERK), total ERK (T-ERK). (C) Target reduction of Drosha and Malat1 RNAs was quantified by qRT-PCR analysis of A431 cells treated with TGF or TGF and PD174265. Data are relative to no PS-ASO control. The error bars represent standard deviations from three independent experiments. Activity curves were fitted from data plotted in panels and IC₅₀ was calculated based on a non-liner regression model. P (in red) <0.01, TGF treatment versus control. P values were computed by One-way ANOVA as the concentrations of PS-ASOs were set as random effect. (D) Western analyses of proteins from A431 cells pre-treated with or without PD174265 followed by the treatment of TGF, as in (B).