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 , b FOXA2 expression profiles in hFOB1.19, MNNG-HOS, 143B, U-2OS and Saos-2 were determined using western blot analysis and qRT-PCR. c FOXA2 expression patterns in OS tissues were exploredusing IHC staining. Scale bar = 50 μm. d The scores for IHC images of OS tissues in T1 ( n = 10) and T2 ( n = 30) stages using anti-FOXA2 antibody were shown. e R2 database analysis of 88 OS cases revealed a correlation between FOXA2 expression and both overall and metastasis-free survival rates. Significance levels were set as follows: ns p > 0.05, * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: FOXA2 shRNA plasmids together with their negative controls were sourced from GeneChem, China.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Immunohistochemistry

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 Knockdown efficiency of FOXA2-specific shRNA in MNNG-HOS and 143B cells was determined using western blot. b CCK-8 experiments demonstrated that FOXA2 silencing reduced MNNG-HOS and 143B cell proliferation. c , d Colony formation assays showed that FOXA2 knockdown inhibited OS cell (143B and MNNG-HOS) proliferation. e , f The migration and invasion capabilities of cells were evaluated using Transwell assays. Data were expressed as mean ± SD. Scale bar = 100 μm. Significance levels were set as follows: ns p > 0.05, * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: FOXA2 shRNA plasmids together with their negative controls were sourced from GeneChem, China.

Techniques: Knockdown, shRNA, Western Blot, CCK-8 Assay, Migration

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 TEM images of mitochondria.Scale bar = 5 μm. b Glutathione assay. c MDA assay. d Lipid ROS production in MNNG-HOS and 143B cells was assessed using C11-BODIPY581/591 staining following transfection with sh-FOXA2 plasmid. Scale bar = 200 µm. e Western blot analysis of GPX4 and xCT protein levels in FOXA2 knockdown cells and negative controls. 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: Glutathione Assay, Multiple Displacement Amplification, Staining, Transfection, Plasmid Preparation, Western Blot, Knockdown

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

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 TEM images of mitochondria. Scale bar = 5 μm. b Glutathione assay. c MDA assay. d Lipid ROS production was assessed in 143B and MNNG-HOS cells using C11-BODIPY581/591 staining across the FOXA2 knockdown, Rheb-overexpressing FOXA2 knockdown and negative control groups. Scale bar = 200 µm. 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: Glutathione Assay, Multiple Displacement Amplification, Staining, Knockdown, Negative Control

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 CCK-8 experimental outcomes in 143B and MNNG-HOS cells for the FOXA2 knockdown, Rheb-overexpressing FOXA2 knockdown, and negative control groups. b , c Colony formation assay outcomes in 143B and MNNG-HOS cells for the FOXA2 knockdown, Rheb-overexpressing FOXA2 knockdown, and negative control groups. d , e Cell migration and invasion capabilities were evaluated using Transwell assays. 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: CCK-8 Assay, Knockdown, Negative Control, Colony Assay, Migration

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 , b Morphological features of xenograft tumors in sh-FOXA2, sh-FOXA2 + Rheb, and sh-Control groups ( n = 5 for each group). Scale bar = 1 cm. Tumor measurements were conducted at five-day intervals. c Tumor weight in sh-FOXA2, sh-FOXA2 + Rheb, and sh-Control groups. d Tumor volume in sh-FOXA2, sh-FOXA2 + Rheb, and sh-Control groups. ( e , f ) Representative images of Ki67-stained xenograft tumors obtained in nude mice in every experimental group. Scale bar = 50 μm. Quantification of positive cells. 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: Control, Staining