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Transcriptional and translational changes of GzmA, GzmB and Perforin in NK92 cells after 24 hours stimulation.

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posted on 20.02.2013, 07:45 by Evelyn Böttger, Gabriele Multhoff, Jürgen F. J. Kun, Meral Esen

NK92 cells were left untreated (ns) or cultured with IL-12/-18 (IL), IFN-α, uRBC or iRBC (1∶3) for 24 h. A: Changes on transcriptional level of stimulated cells in comparison to untreated cells were analyzed for GzmA, GzmB and Perforin. β-Actin expression served as house-keeping gene normalizer. Data are represented as mean ± SD (student's t test * p<0.05; ** p<0.01) and are representative of three independent experiments each performed in duplicate. B: After 24 h, 1×107 NK92 cells were lysed, incubated 30 min on ice and subsequently centrifuged at 13000× g for 15 min at 4°C. 10 µg of total supernatant protein were separated by 7.5% SDS-PAGE and blotted onto a nitrocellulose membrane. After blocking, membranes were incubated for 1 h with anti-β-Actin (lane 1), anti-hGzmA (lane 2), anti-hGzmB (lane 3), or anti-hPrf (lane 4).