Western analysis of EphB4 in prostate cancer cell lines. Western analysis of 20 μg EphB4 protein in prostate cancer cell lines LNCaP (L), DU145 (D) and PC3 (P). (A) The immunoblot was incubated sequentially with antibodies specific to EphB4, α-actin and calnexin and exposed to autoradiographic film for the indicated times. MCF10A engineered to express EphB4 (5 μg protein lysate) was used as a positive control. The arrow indicates the expected size of each protein – 120 kDa for EphB4, 43.2 kDa for α-actin and 90 kDa for calnexin. The size of the marker proteins is shown on the left in kDa. (B) Coomassie stained duplicate gel for loading comparison.