Sorbitol Destroyed Intestinal Microfold Cells (M Cells) Development through Inhibition of PDE4-Mediated RANKL Expression

Figure 3

CREB is critical for sorbitol-mediated RANKL expression: (a) reporter gene containing RANKL promoter was transfected into CaCO₂ cells combined with pGL4.74 plasmid for 24 hr, CaCO₂ cell was treated with 100 mM mannitol and 100 mM sorbitol for further 24 hr, the relative luciferase unit was detected using dual-luciferase reporter assay system. Data presented as the mean ± s.e.m. of three independent experiments and were analyzed by two ANOVA, , ; (b) after serum-free for 24 hr, CaCO₂ cells were stimulated for 1 hr, and the whole cell was fixed and lysed to incubate with an anti-CREB antibody, the chromosome fragments binding to CREB were amplified and quantified by real-time PCR with RANKL promoter primer. Data presented as the mean ± s.e.m. of three independent experiments and were analyzed by two ANOVA, ; (c) ELISA was employed to detect RANKL level in indicated group in CaCO₂ cells, data presented as the mean ± s.e.m. of five independent experiments and were analyzed by one ANOVA, .

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(b)

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