FHC silencing in K562 cells reduces proliferation rate via RAF1/MAPK pathway inhibition and is associated with <i>c-Myc</i>down-regulation.

<p>A) Real-time PCR of <i>RAF1</i> mRNA performed on K562 shRNA, K562 shFHC and K562^shFHC/pc3FHC. Results are representative of two different experiments B) Western Blot analysis for pERK1/2 was performed on 50μg of total protein extract from K562shRNA and K562shFHC cells. Total ERK1/2 was used as loading control. Results are representative of three different experiments. C) Western Blot analysis for pERK1/2 and FHC was performed on 50μg of total protein extract from K562shRNA and K562shRNA+FHC. Total ERK1/2 and γ-Tubulin were used as loading controls. Results are representative of two different experiments. D) Equal number of starved silenced and un-silenced cells were plated into a 96-well plate, incubated for 72 h and analysed by MTT assay. Proliferation of FHC-silenced cells is reduced of about 35% compared to controls. Data are presented as mean ±standard deviation. E) Real-time PCR of c-<i>Myc</i> mRNA performed on K562 shRNA, K562 shFHC and K562^shFHC/pc3FHC. Results are representative of two different experiments.