Induction of LMP-1 expression by IL-4 and IL-13 in the KMH2-EBV cells. (A) Expression of EBNA-2, LMP-1, and β-actin in total cell lysates of KMH2-EBV treated for 24 hours with control L cells or CD40L-L cells, with or without IL-4 (50 ng/mL), assessed by Western blotting. In all of the immunoblots, the arrow indicates the expression of the lower molecular weight LMP-1 (“truncated” LMP-1). (B) LMP-1 expression after activation of KMH2-EBV cells with IL-4 (50 ng/mL) for the indicated times. (C) Expression of LMP-1 in the KMH2-EBV cells treated with the indicated cytokines (50 ng/mL) for 24 hours. (D) Expression of LMP-1, EBNA-2, and β-actin in the KMH2-EBV cells after treatment with the indicated doses of IL-4 or IL-13 for 24 hours. (E) Immunofluorescence staining for LMP-1 or EBNA-2 of the IL-4– or IL-13–treated KMH2-EBV cells (50 ng/mL, 24 hours). Nuclei were visualized with Hoechst 33258. Images were generated with a Leitz DM RB microscope (Leica Microsystems) using a 63×/1.32 NA oil immersion objective lens. Images were captured with a Hamamatsu dual-mode cooled charged coupled device camera (C4880) and Hipic 6.4.0 software (Hamamatsu Photonics Deutschland). Pictures were edited for opitmal color contrast with Adobe Photoshop 7 (Adobe Systems). (F) Induction of LMP-1 in KMH2-EBV(PM) cell line by the indicated doses of IL-4 or IL-13 after 24 hours of treatment. (G) Expression of LMP-1 in KMH2-EBV cells treated with IL-4 (50 ng/mL) for 6, 12, 24, 48, or 72 hours. After 72 hours the cells were washed in phosphate-buffered saline and replated for an additional 24, 48, or 72 hours in complete RPMI medium without IL-4.