Mouse ifn λ2

With exception of Fludarabine, obtained from Tocris Biosciences and CCCP obtained from Sigma, all inhibitors were obtained from Calbiochem. Cells were incubated with 10–50 μM Fludarabine, 2 μM Pyridone 6, 10–50 μM AG490, 10–100 μM 1,2,3,4,5,6-Hexabromocyclohexane (JAK2 inhibitor II, HBC), 5 μM Bay11, 50 μM PDTC, 30 μM PD98059, 100 μM SP600125, 10 μM SB202190 for 30 min, prior to infection or treatment. In case of IFN treatment, cells were incubated in fresh medium with 0.01 μg/ml human IFN-λ1 or mouse IFN-λ2 (Peprotech), 10 U/ml human IFN-β or 50 U/ml mouse IFN-β (PFL Interferon Source) for 2–5h. Keratinocytes were incubated in media containing either 10 μM CCCP or DMSO. Cells were incubated for 48 hours and then infected or incubated with 250 nM MitoTracker Deep Red FM (Molecular Probes) for 30 min at 37°C prior to fixation in 2% paraformaldehyde in PBS. Confocal images were acquired using a spinning disk confocal head (CSU-X1, Perkin Elmer Co., Boston, MA) coupled to a fully-motorized inverted Zeiss Axiovert 200M microscope equipped with a 63X lens (Pan Apochromat, 1.4 NA). The imaging system operates under control of SlideBook 5 (Intelligent Imaging Innovations Inc, Denver, CO). Micrographs were processed with Adobe Photoshop.