Figure 9.
Overexpression of EGFR increases PS-ASO activity in HEK cells. (A) Western analyses for proteins in control or EGFR overexpressing HEK cells treated with EGF for indicated time. The blot was probed sequentially with different antibodies detecting phosphorylated EGFR (P-EGFR), total EGFR (T-EGFR) and total ERK (T-ERK) as loading control. (B) Representative images of immunofluorescent staining for EGFR (green) in HEK cells overexpressing EGFR. Nuclei were stained with Hoechst 33342 (blue). Scale bar, 5 μm. (C) Reduction of Malat1 RNA was quantified by qRT-PCR in HEK cells with or without overexpression of EGFR. Data are relative to untreated cells. Activity curves were fitted from data plotted in panels based on a non-linear regression model. The error bars represent standard deviations from 3 independent experiments. P< 0.01 (in red), HEK versus HEK (O.V. EGFR); P value was computed by One-way ANOVA as the concentrations of PS-ASOs were set as random effect; IC50 was calculated based on a non-liner regression model. (D) Intracellular fluorescence of Cy3-PS-ASO was quantified by flow cytometry to determine PS-ASO uptake (RFU) in HEK cells with or without overexpression of EGFR.