Figure 6

From: p400 function is required for the adenovirus E1A-mediated suppression of EGFR and tumour cell killing

Figure 6

(a) Western blot analysis of HSC3 HI and p400 mixed populations. p400 was detected using anti-p400 RW144 monoclonal primary antibody and tubulin was detected using anti-tubulin monoclonal antibody (Sigma). The intensity of tubulin and p400 bands from the western blot was measured using Version 2.0 of Aida 2D Densitometry software. The amount of p400 protein was normalized with tubulin expression level to correct for loading differences. The percentage of p400 expression of HSC3-p400mp2 was calculated with respect to HSC3-HImp1, which was set at 100%. (b) MTT analysis of HSC3 HI and p400 mixed populations 48 and 72 h after infection with Ad-Del and Ad-E1A with a MOI of 10. Error bars indicate s.d. Statistical analysis was performed using Tukey's Multiple Comparison Test, P-values are shown. (c) Western blot analysis of HSC3-HImp1 and HSC3-p400mp2 48 and 72 h after infection with indicated recombinant adenoviruses. Cells were lysed at 48 and 72 h post-infection, equal amounts of total protein from each sample was separated by SDS–PAGE, transferred to nitrocellulose and hybridized to different antibodies as described in the Materials and methods section. The percentage of EGFR expression of cells infected with Ad-E1A was calculated with respect to the EGFR expression of control Ad-Del-infected cells, which was set at 100%. Abbreviations: EGFR, epidermal growth factor receptor; MTT, 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide; SDS–PAGE, lauryl sulphate–polyacrylamide gel electrophoresis.