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. 2008 Oct;125(2):229–240. doi: 10.1111/j.1365-2567.2008.02833.x

Figure 4.

Figure 4

Simultaneous electroporation of a mixture of mRNAs encoding CD80, 4-1BBL, OX40L, IL-12p35, IL-12p40 and actin into activated B cells. (a) B cells activated for 2 days with a combination of CpG and anti-CD40 were transfected with either actin mRNA (40 μg/ml) or a mixture of IL-12p35 and IL-12p40 mRNAs at various concentrations (20 μg/ml, 40 μg/ml or 80 μg/ml each). Culture supernatants were collected 24 hr after transfection and assayed by IL-12p70 enzyme-linked immunosorbent assay (ELISA). (b) Activated B cells were transfected with either a mixture of IL-12p35 and IL-12p40 mRNAs or a mixture of IL-12p35, IL-12p40, CD80, OX40L, 4-1BBL and actin mRNAs (each of them at 25 μg/ml). At 24 hr after transfection, production of IL-12p70 was measured by ELISA and expression of CD80, 4-1BBL and OX40L was analysed by flow cytometry. The numbers in the histograms are percentage of positive cells after transfection with a mixture of mRNAs encoding IL-12p35, IL-12p40, CD80, OX40L, 4-1BBL and actin and, in parentheses, percentage of positive cells after transfection with mixture of mRNAs encoding IL-12p35 and IL-12p40. (c) Activated B cells were transfected with either actin mRNAs (150 μg/ml; grey line) or a mixture of IL-12p35, IL-12p40, CD80, OX40L, 4-1BBL and actin (25 μg/ml each; black line). Amounts of CD80, 4-1BBL and OX40L were determined by flow cytometry at the indicated times after transfections. IL-12p70 production by the cells during the indicated culture periods after transfection was measured by ELISA. The data, except the histogram, are means of three experiments and the histograms are a representative of three experiments. Error bars represent SD.