Article Title: Protein Kinase C Signaling in Adenoviral Infection
Figure Lengend Snippet: Activation of cSrc and caveolin-1 in endosomes occurs downstream of PKC α . (A) Western blot analysis of endosomal preparations of serum starved corneal fibroblasts mock infected (M), DMSO (V), or calphostin C pretreated and virus infected (Cal.C+V) for two hr. Postnuclear extracts were subjected to endosome purification using sucrose gradient centrifugation. After SDS-PAGE, membranes were blotted with antibody to p-PKC α , then stripped and reprobed with antibody against phosphorylated cSrc (p-cSrc), and then stripped and reprobed with antibody to phosphorylated caveolin-1 (p-Cav-1). Virus infection increased expression of all three proteins, and all were markedly reduced by pretreatment with calphostin C. (B) cSrc kinase assay performed at 2 h post infection on pooled endosomal preparation fractions 7 through 10 shows increased cSrc activity in virus infection (V) than in mock infection (M) or in virus infected cells pretreated with calphostin C (Cal.C + V, * p
Article Snippet: Antibodies to total cSrc and phospho-caveolin-1 were purchased from EMD Millipore (Billerica, MA) and BD Bioscience (San Jose, CA).
Techniques: Activation Assay, Western Blot, Infection, Purification, Gradient Centrifugation, SDS Page, Expressing, Kinase Assay, Activity Assay