FOXO1 Mediates Advanced Glycation End Products Induced Mouse Osteocyte-Like MLO-Y4 Cell Apoptosis and Dysfunctions

Figure 5

FOXO1 directly binds to the caspase-3, sclerostin, and RANKL promoter regions. ChIP assays were performed in MLO-Y4 cells with pulldown by FOXO1 antibody and PCR amplification of a region of the caspase-3, sclerostin, and RANKL promoter flanking the FOXO1 consensus response element. Results were compared to both its relative control IgG and the FOXO1 binding of the BSA control group. (a) Quantifications of caspase-3 ChIP assay. (b) Quantifications of sclerostin ChIP assay. (c) Quantifications of RANKL ChIP assay. MLO-Y4 cells were cotransfected with empty vector or FOXO1 vector and caspase-3, sclerostin, and RANKL luciferase reporter and Renilla control construct. Results were compared to control (empty vector) and FOXO1 expression plasmid in the BSA control group. (d) Quantifications of caspase-3 luciferase assay. (e) Quantifications of sclerostin luciferase assay. (f) Quantifications of RANKL luciferase assay. All error bars are standard deviation of the mean. ^# compared to BSA control group. ^## compared to the BSA FOXO1 group.

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