Journal: International Journal of Molecular Sciences

Article Title: Cytokine-Modulated Natural Killer Cells Differentially Regulate the Activity of the Hepatitis C Virus

doi: 10.3390/ijms19092771

Figure Lengend Snippet: Effect of recombinant IL-21 on expression of CD56, STAT proteins, and IFN-γ in NK-92 cells. NK-92 cells were cultured in the presence or absence of recombinant IL-21 (0.2 ng/mL) for 6 h and stained with fluorescent anti-CD56 antibodies. ( A ) Representative FACS plots showing CD56 expression of NK-92; ( B ) % CD56 +dim NK-92. Alternatively, NK-92 cells were incubated with or without recombinant IL-21 (0.2 ng/mL) for 6 h and subsequently cocultured with J6/JFH-1-huh 7.5 cells or naïve huh 7.5 cells for 12 h. Cell lysates from NK-92 cells only were assessed for the expressions of STAT1 and STAT5 proteins. β-actin was served as the loading control. IFN-γ production (pg/mL) in culture supernatants was determined by ELISA. ( C ) Expressions of STAT1 and STAT5 proteins by Western blot; ( D ) Relative band intensity of STAT1 and STAT5 proteins compared with the loading control from at least three independent experiments; ( E ) IFN-γ production (pg/mL).

Article Snippet: Cell free supernatants were harvested to measure the production of IFN-γ using a human enzyme linked immunosorbent assay kit (BD Biosciences).

Techniques: Recombinant, Expressing, Cell Culture, Staining, Incubation, Enzyme-linked Immunosorbent Assay, Western Blot