Replication Data for: FOXO3a promotes odontoblast differentiation in human dental pulp cells in coordination With RUNX2

This dataset comprises supplementary figures that support the experimental validation and quality control procedures associated with this study investigating the role of Forkhead box O3a (FOXO3a) in odontoblast differentiation of human dental pulp cells (HDPCs). Specifically, the dataset includes: (1) cell viability analyses performed to determine the appropriate working concentrations of carbenoxolone (CBX), a FOXO inhibitor, and SC79, an AKT activator, prior to functional experiments; (2) cell viability assessments under luciferase reporter assay transfection conditions to verify that plasmid transfection and pharmacological treatments did not influence cell viability or confound reporter activity; and (3) evaluation of transfection efficiency using an EGFP-expressing control plasmid under the same conditions employed for FOXO3a overexpression in HDPCs. Supplementary Figure S1. Effects of carbenoxolone and SC79 on HDPC viability. HDPCs were treated with increasing concentrations of carbenoxolone (CBX; 0, 25, 50, 100, 150, and 200 μM) or SC79 (1, 2.5, and 5 μM) for 24 h, and cell viability was evaluated using the WST-8 assay to determine the appropriate working concentrations for subsequent functional experiments. Supplementary Figure S2. Cell viability under luciferase reporter assay transfection conditions. HDPCs were transfected with DSPP (pGL3-DSPP) or RUNX2 (pGL3-RUNX2) luciferase reporter constructs together with FOXO3a and/or RUNX2 expression plasmids, with or without 150 μM CBX treatment. Cell viability was assessed before the luciferase assay to confirm that reporter activity was not influenced by cytotoxicity or differences in cell number. Supplementary Figure S3. Verification of transfection efficiency in HDPCs. HDPCs were transfected with an EGFP-expressing control plasmid under the same Lipofectamine LTX-mediated transfection conditions used for FOXO3a overexpression. Transfection efficiency was determined by quantifying the percentage of EGFP-positive cells among DAPI-stained nuclei.