Quantification of Viral and Cellular Proteins

The isolation of nuclear and cytoplasmic fractions was performed as reported previously^{60 (link)}. Proteins derived from nuclear and cytoplasmic fractions were quantified by ‘DC Protein Assay’ (Bio-Rad), and equal amounts were used for western blot analysis to evaluate the accumulation of both viral and cellular proteins. Resolved proteins by SDS-PAGE were transferred to nitrocellulose membranes (Biorad). The membranes were incubated overnight at 4 °C with the appropriate primary antibody and successively probed with secondary antibodies. Quantitative densitometry analysis of immunoblot band intensities was performed by using the TINA software (version 2.10, Raytest, Straubenhardt, Germany).

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Venuti A., Musarra-Pizzo M., Pennisi R., Tankov S., Medici M.A., Mastino A., Rebane A, & Sciortino M.T. (2019). HSV-1\EGFP stimulates miR-146a expression in a NF-κB-dependent manner in monocytic THP-1 cells. Scientific Reports, 9, 5157.

Publication 2019

DC Protein Assay nitrocellulose membranes TINA software

Antibodies Antibody Assay Cellular Cytoplasmic Densitometry Immunoblot analysis Isolation Membranes Nitrocellulose Proteins Sds page Tina Western blot analysis

Corresponding Organization : University of Messina

Other organizations : University of Tartu

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Variable analysis

independent variables

Nuclear and cytoplasmic fractions

dependent variables

Accumulation of both viral and cellular proteins

control variables

Equal amounts of proteins derived from nuclear and cytoplasmic fractions were used for western blot analysis

controls

No positive or negative controls were explicitly mentioned in the provided information.

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