The Nod2 Agonist Muramyl Dipeptide Cooperates with the TLR4 Agonist Lipopolysaccharide to Enhance IgG2b Production in Mouse B Cells

Figure 6

Effects of LPS and MDP on cell viability, cell proliferation, IgG2b production, and germline γ2b transcripts expression in Rip2-deficient B cells. Resting B cells were purified from WT and Rip2-deficient (Rip2^-/-) B cells and stimulated with MDP (10 μg/mL) and LPS (1 μg/mL). (a) After 2 and 3 days of culture, cell viability (OD) and proliferation were measured by EZ-Cytox assay and CFSE assay, respectively. Low CFSE intensity cell (%) means the proportion of proliferating cells. (b) After 7 days of culture, supernatants were harvested and the levels of Ab production were measured using isotype-specific ELISA. Data shown are averages of triplicate cultures with SEM error bars. SEM: standard error of the mean. (c) After 2.5 days of culture, RNAs were isolated and the levels of germline γ2b transcripts were measured by RT-PCR. The levels of germline γ2b transcripts were measured by semiquantitative RT-PCR with 1/5 and 1/25 diluted cDNA (c, lower panel).

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