Fig 2. FHC over-expression reduces cell proliferation.

(A) Western blot analysis for FHC was performed on 50μg of total proteins extracted from H460 over-expressing FHC (H460^pcFHC) and H460 control cells (H460^pcDNA). γ-Tubulin was used as a loading control. Blots are representative of three independent experiments. The graph representsthe mean of the optical densities (*p < 0.001 compared with control cells). (B) Real-time PCR analysis of FHC mRNA levels was performed on total RNA from H460^pcFHC and H460^pcDNA. Results are representative of three different experiments (* p<0.05 compared with H460^pcDNA). (C) Cell proliferation was assessed using the MTT method as indicated in the Materials and Methods section. Final results represent mean ± SD of three independent experiments each performed in octuplicate (*p< 0.05 compared with H460^pcDNA). (D) Western blot analysis for CCND1, p53 and pAKT were performed on 50 μg of total proteins extracted from H460^pcFHC and H460^pcDNA. Blots are representative of three independent experiments. γ-Tubulin and AKT were used as loading controls.