Figure 4.

Inhibition of EGFR by RGFP966 is associated with HDAC3. (A), Huh7 cells were treated with indicated doses of RGFP966 (RGFP), or vehicle. After 40 hrs, cells were harvested, and total RNA was extracted. Relative expression of EGFR mRNA was determined by real-time PCR. GAPDH was used as internal control. Data are represented as mean ± SD from three independent experiments. P value refers to two-sided t test. (B), Huh7 cells were separately transfected with empty vector and expression vector for HA-tagged HDAC3. After 40 hrs, cells were harvested, and total RNA was extracted. Relative expression of EGFR mRNA was determined by real-time PCR. GAPDH was used as internal control. Data are represented as mean ± SD from three independent experiments. P value refers to two-sided t test. (C), Huh7 cells were separately transfected with empty vector and different doses of HA-tagged HDAC3 expression plasmids. After 48 hrs, cells were harvested, and Western Blot analysis was performed with the indicated antibodies. GAPDH was used as internal control. (D), Huh7 cells were separately transfected with empty vector and expression vector for HA-tagged HDAC3. After 24 hrs, 2.5 x10⁴ cells were plated into a transwell chamber. After 40 hrs, the invaded cells were stained, and representative images were photographed. (E), Huh7 cells were treated with indicated doses of RGFP966 (RGFP), or vehicle. After 48 hrs, cells were harvested and Western Blot analysis was performed with the indicated antibodies. GAPDH was used as internal control. (F), Huh7 cells were separately transfected with empty vector and expression plasmids for HA-HDAC3, and then were treated with RGFP966 (RGFP, 10μM) and vehicle for 48hrs. Then, cells were harvested, and Western Blot analysis was performed with the indicated antibodies. GAPDH was used as internal control.