Figure 1

HAdV-19 infection of keratocytes induces activation of p38 MAPK. Lysates were prepared 30 min after HAdV-19 or mock infection. (A) Western blot analysis using antibodies against phospho- and total p38 MAPK reveals phosphorylation in HAdV-19 infected cells, reduced in the presence of the Src inhibitor PP2 (10 μM). Densitometry values for the phosphorylated protein (normalized to the corresponding total protein) are shown above each lane. (B) An in vitro p38 MAPK assay performed at 15 and 30 min after infection shows increased phosphorylation of the ATF-2 substrate signifying p38 MAPK activity upon HAdV-19 infection of keratocytes. (C) Densitometric quantification from three experiments of the phosphorylated ATF-2 band in the p38 MAPK assay revealed increased activity in virus infected keratocytes at both 15 and 30 min post-infection (p = 0.0012 and p = 0.0025, respectively). (D & E) Western blot analysis using phospho- and total antibodies against HSP27 and ATF-2 respectively, reveals increases in phosphorylation in HAdV-19 infected cells that was absent in the presence of PP2.