Figure 1.
EBNA2 and ICOSL expression in B-lymphoma cell lines infected with EBV. (A) Immunoblots show EBNA2 expression in parental, B95.8, and P3HR1 virus–converted RAMOS (E95D and EHRB RAMOS, respectively) cells and BJAB, BJAB-B95.8, and BJAB-EHR1 cells. EBNA2 expression in U2932 DLBCL cell line infected with the recombinant EBVGFP viral strain is also shown. U2932 EBVGFP cl.A is negative for EBNA2, whereas U2932 EBVGFP cl.B is positive. Total cell lysates were electrophoresed, and EBNA2 expression was verified by using anti-EBNA2 (PE2) monoclonal antibodies. The housekeeping protein, β-actin, was used as loading control. Densitometry analysis of immunoblots is shown in supplemental Figure 1A. (Bi) ICOSL expression in B and P virus–converted B-lymphoma cell lines, P3HR1 and Jijoye pair, and EREB2-5–carrying inducible EBNA2. Mean fluorescent intensity (MFI) was measured by flow cytometer, CytoFLEX. One of 3 representative experiments is shown. Phycoerythrin (PE)-conjugated ICOSL antibodies were used for the experiments. (Bii) Histograms show the average ICOSL MFI of 3 experiments. The significance of ICOSL MFI between EBNA2− vs EBNA2+ cell lines was calculated using 2-tailed unpaired t test; ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ∗∗∗∗P < .0001.

EBNA2 and ICOSL expression in B-lymphoma cell lines infected with EBV. (A) Immunoblots show EBNA2 expression in parental, B95.8, and P3HR1 virus–converted RAMOS (E95D and EHRB RAMOS, respectively) cells and BJAB, BJAB-B95.8, and BJAB-EHR1 cells. EBNA2 expression in U2932 DLBCL cell line infected with the recombinant EBVGFP viral strain is also shown. U2932 EBVGFP cl.A is negative for EBNA2, whereas U2932 EBVGFP cl.B is positive. Total cell lysates were electrophoresed, and EBNA2 expression was verified by using anti-EBNA2 (PE2) monoclonal antibodies. The housekeeping protein, β-actin, was used as loading control. Densitometry analysis of immunoblots is shown in supplemental Figure 1A. (Bi) ICOSL expression in B and P virus–converted B-lymphoma cell lines, P3HR1 and Jijoye pair, and EREB2-5–carrying inducible EBNA2. Mean fluorescent intensity (MFI) was measured by flow cytometer, CytoFLEX. One of 3 representative experiments is shown. Phycoerythrin (PE)-conjugated ICOSL antibodies were used for the experiments. (Bii) Histograms show the average ICOSL MFI of 3 experiments. The significance of ICOSL MFI between EBNA2− vs EBNA2+ cell lines was calculated using 2-tailed unpaired t test; ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ∗∗∗∗P < .0001.

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