Primary antibodies specific for adenoviral proteins used in this study were E2A-72K mouse mAb B6-8 [29 (link)] and L4-100 K rat mAb 6B10 [30 (link)]. Cellular protein-specific primary antibodies included polyclonal rabbit Ab raised against the PML protein (Abcam, ab72137) and ß-actin mouse mAb AC-15 (Sigma-Aldrich, Inc.). The used secondary antibodies conjugated to horseradish peroxidase (HRP) for detection by immunoblotting were anti-rabbit IgG, anti-rat IgG and anti-mouse IgG (Jackson/Dianova).
After cell harvesting 24 h p.i., protein extracts were prepared in RIPA lysis buffer, as described elsewhere [31 (link)] and separated by SDS-PAGE. Samples were transferred to nitrocellulose blotting membranes (0.45 µm) and proteins were visualized by immunoblotting. Fiji [32 (link)] was used for quantification analysis and amounts of proteins were calculated relative to actin.
After cell harvesting 24 h p.i., protein extracts were prepared in RIPA lysis buffer, as described elsewhere [31 (link)] and separated by SDS-PAGE. Samples were transferred to nitrocellulose blotting membranes (0.45 µm) and proteins were visualized by immunoblotting. Fiji [32 (link)] was used for quantification analysis and amounts of proteins were calculated relative to actin.
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