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. 2002 Apr 2;99(7):4514–4519. doi: 10.1073/pnas.072071599

Figure 5.

Figure 5

Antibody production and germ-line CH gene expression in nfkb1−/−c-rel−/− mice. (A) Serum Ig levels in naive mice. Levels of serum Igs (μg/ml) in naive 8- to 10-week-old litter-matched mice was determined by isotype-specific ELISA. Filled, hatched, empty, and stippled bars correspond to wild-type, nfkb1−/−, c-rel−/−, and nfkb1−/c-rel−/− mice, respectively. Five mice of each genotype were examined. IgG1 levels were below the level of detection (ND) in nfkb1−/c-rel−/− mice. (B) Humoral response to NP-KLH. Five age-matched mice of each genotype were immunized with NP-KLH and serum samples were collected after 21 days. The concentration (μg/ml) of NP-specific IgG1 is indicated for wild-type (filled bars), nfkb1−/− (hatched bars), and c-rel−/− (empty bars) mice. Antigen-specific IgG1 was below the levels of detection (ND) in nfkb1−/c-rel−/− mice. (C) Germ-line CH transcript expression. RNA isolated from equal numbers of wild-type (lane 1), nfkb1−/− (lane 2), c-rel−/− (lane 3), and nfkb1−/−c-rel−/− (lane 4), B cells stimulated in culture for 6 h with various mitogen plus cytokine combinations (13) was subjected to semiquantitative reverse transcription–PCR using primers specific for Cγ3, Cγ1, Cɛ, and Cα germ-line RNAs and β-actin mRNA as a control. The PCR products were then fractionated on agarose gels. B cells were stimulated with LPS alone (Cγ3 RNA), LPS plus IL-4 (Cγ1,Cɛ RNA), and LPS plus transforming growth factor β (Cα RNA).