Fig. 3

Detection of GC cell populations and determination of mRNA expression levels of class switch recombination-related genes. Mice were intranasally immunized with PBS, 1.1 μg tFomA (tA), 5.1 μg FlaB-LK1-tFomA (BtA), 4 μg Hgp44 (H), 8 μg Hgp44-LK3-FlaB (HB) or a mixture of 5.1 μg BtA and 8 μg HB (BtA + HB) three times at 2-week interval. Two weeks after the last immunization, the cLNs cells were prepared to evaluate GC cell populations by flow cytometry (a, b), and cytokine mRNA expression levels were determined by qRT-PCR (c). Total RNA was isolated from the cLN cells, and the mRNA expression levels were measured by qRT-PCR for the indicated genes. The data are presented as the mean ± SEM in each group. N = 5, **P < 0.01, ***P < 0.001, NS non-significant