Figure 5
Stimulation or inhibition of EGFR signalling does not affect reovirus cytotoxicity in Cal27 cells. Cells were treated for 1 hr with 200 nM epidermal growth factor (EGF), 400nM anti-EGFR antibody (ICR62), 1 μM Iressa or 100 μM Tyrphostin AG99 (Tyrp), then either lysed, resolved on 8% Precise Protein Gels and probed for total EGFR, phospho-Tyr1068 EGFR and GAPDH or α-tubulin as loading controls, or infected with reovirus at 1.9×109 TCID50/ml and assayed for cell survival by MTT at 96 hours post-infection. Reovirus was diluted as follows: 1:16000 (20%), 1:4000 (50%) and 1:500 (80%). A. EGF stimulation does not increase reoviral cytotoxicity. B, C, D. ICR62-, gefitinib- (Iressa) and Tyrphostin-mediated inhibition of EGFR did not inhibit reoviral cytotoxicity. Means are calculated from 3 independent experiments and error bars represent SEMs. E. Western blot analysis showing effect of EGF, ICR62, Gefitinib (Iressa) and Tyrphostin on EGFR signaling.