Journal: Vaccines

Article Title: E4orf1 Suppresses E1B -Deleted Adenovirus Vaccine-Induced Immune Responses

doi: 10.3390/vaccines10020295

Figure Lengend Snippet: Effects of E4orf1 on innate cytokine gene expression in infected human epithelial cell lines. HeLa cells were infected at a MOI of 50 with Δ E3, Δ E1B and Δ E4 1 Ads and incubated for 48 h as described in Materials and Methods. ( A , B ) Western blotting using whole cell lysate shows the presence or absence of Akt phosphorylation at Ser473 and Thr308, with ( A ) Ad DNA binding protein (DBP) and ( B ) cells treated with or without AktIV. PCR for Ad fiber is used as the Ad infection control. ( C ) The fold change in expression levels of all the cytokines evaluated in HeLa cells are plotted. ( D , E ) The magnitudes of the induced/upregulated cytokines in each group are compared in ( D ) HeLa cells and ( E ) A549 cells. The differences in mean values are highlighted with colored boxes. For each cell line n = 2 biological repeats performed in triplicate. Mean plus SEM are plotted. p values obtained by Mann–Whitney unpaired t -test and Wilcoxon matched-pairs signed-rank test * = p < 0.05, ** = p < 0.01.

Article Snippet: We used the human cervical carcinoma-derived HeLa cell line (ATCC ® CCL-2TM) because most of what is known about Ad is in the context of this cell line.

Techniques: Gene Expression, Infection, Incubation, Western Blot, Phospho-proteomics, Binding Assay, Control, Expressing, MANN-WHITNEY