Figure 6.

a. MetR cells have increased expression of EMT markers: 50ug of total protein extracted from MetR and LNCaP cells were loaded on SDS page gels. Western blots ere performed using anti-vimentin, anti-twist and anti-GAPDH antibodies. b. Axl overexpression in LNCaP cells caused an increase in expression of EMT proteins: Cells were either transfected with empty vector or with pcDNA-Axl plasmid. Total protein was extracted and western blotting was repeated as above to detect the protein levels in LNCaP and LNCaP-Axl cells. c, d. EMT markers are down-regulated when Axl is knocked down in Du145 cells: Axl was stably knockdown in Du145 cells using lentiviral transduction. Total RNA was extracted from Du145 and Du145-sh-Axl cells and RT-PCR was done with snail, slug, twist, E-cadherin and GAPDH primers. d. 50ug of total protein from Du145 and Du145-sh-Axl cells was loaded on a SDS page gel and western blotting was done using Axl, twist and GAPDH antibodies.