Journal: World Journal of Surgical Oncology
Article Title: FOXA2 promotes the occurrence and development of osteosarcoma cells by regulating ferroptosis through the Rheb/mTOR axis
doi: 10.1186/s12957-026-04308-4
Figure Lengend Snippet: a Nine common target genes were identified by overlapping the mTOR signaling pathway gene set with FOXA2 target genes predicted by CHIP-ATLAS, ENCODE, GTRD, and hTFtarget databases. b qRT-PCR analysis of EIF3A, EIF4A1, EIF4EBP1, mTOR, PI3KCA, Rheb, TSC1, RPS6KB1, and TSC2 in MNNG-HOS and 143B cells. c JASPAR database predicted binding sites between transcription factor FOXA2 and Rheb promoter. d ChIP assay was conducted to verify the FOXA2-binding site in the Rheb promoter region within 143B and MNNG-HOS cell lines. Controls comprised IgG and input fractions. e Luciferase reporter plasmids with either wild-type or mutant Rheb2 promoters were used for assessment of luciferase activities in 143B and MNNG-HOS. The mutation site is highlighted in red. f 143B and MNNG-HOS cells with FOXA2 knockdown, FOXA2 knockdown combined with Rheb overexpression, and negative controls were used to analyze Rheb, total and phosphorylated mTOR, xCT, and GPX4 at the protein level. Data are presented as mean ± SD. Significance levels were set as follows: ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001
Article Snippet: FOXA2 shRNA plasmids together with their negative controls were sourced from GeneChem, China.
Techniques: Quantitative RT-PCR, Binding Assay, Luciferase, Mutagenesis, Knockdown, Over Expression