File S1 - The Differential Expression of EphB2 and EphB4 Receptor Kinases in Normal Bladder and in Transitional Cell Carcinoma of the Bladder

Figure S1 & S2 and Table S1 & S2. Figure S1. Representative EphB4/EphB2 immunostaining on tumor and normal urothelium obtained from the same patient during cystectomy. 2 paired cases are shown. Nuclei were counter-stained with DAPI. Figure S2. EphB4 siRNA knockdown induces apoptosis in bladder cancer cell but not normal bladder cell. 5637 and PD071 cells were grown in 6-well plate and transfected with 50 nM EphB2 or EphB4 siRNA with Lipofectamine 2000 (Invitrogen). 48 hours later, cells were harvested with Cell Dissociation Buffer (Sigma), stained with Annexin V apoptosis quantitation kit (Biotium), and analyzed on flow cytometer LSRII (BD Biosciences). Dead cells were excluded by 7-AAD staining. Data were analyzed with FlowJo (Tree Star). A, EphB2 siRNA had no effect on apoptosis of 5637, whereas EphB4 siRNA led to significant apoptosis. B, Both EphB2 and EphB4 siRNA had no effect on PD071 apoptosis. (PPTX)