Fig 1: The expression levels of three hypoxia-related genes between osteosarcoma cell lines and osteoblasts. (A) Western blotting of the expressions of RASGRP2, KCNJ3, and ACTG2 in hFOB, U20S, and 143B groups. GAPDH serves as an internal standard. The gels have been run under the same experimental conditions. (B) A histogram of the OD values of RASGRP2, KCNJ3, and ACTG2 in each group (n = 3 per group). The obtained data are represented as mean ± SE. Significance: ***p-value < .001, vs. hFOB group.
Fig 2: Immune infiltration analysis of the five gene signatures. (A) Correlation analysis comparing the five prognostic genes and six main immune cells. (B) Correlation analysis of the risk score and 10 subtypes of immune cells. *P<0.05 and **P<0.01. ACTG2, actin γ2; APOD, apolipoprotein D; COPG2IT1, coatomer protein complex subunit γ2 imprinted transcript 1; COL6A6, collagen type VI α6 chain; OMD, osteomodulin; NK, natural killer.
Fig 3: External validation of the prognostic model using TCGA dataset. (A) Distribution of risk scores in TCGA dataset using the five metastasis-related genes from the prognosis prediction model. (B) Survival status of patients with adult soft-tissue sarcoma in TCGA dataset categorized into high- and low-risk score groups. (C) Heatmap showing expression levels of the five prognostic genes stratified by risk score. (D) Kaplan-Meier survival curves for patients divided into the two risk score groups. (E) Receiver operating characteristic curves for predicting overall survival over a period of 1 to 5 years based on the five-gene signature. ACTG2, actin γ2; APOD, apolipoprotein D; COPG2IT1, coatomer protein complex subunit γ2 imprinted transcript 1; COL6A6, collagen type VI α6 chain; OMD, osteomodulin; TCGA, The Cancer Genome Atlas.
Fig 4: Assessment of the prognosis prediction model. (A) Distribution of patients categorized by risk score. (B) Distribution of survival rates associated with risk scores. (C) Heatmap of expression levels for the five prognostic genes stratified by risk score. (D) Kaplan-Meier survival curves for patients grouped according to the two risk score categories. (E) ROC curves for predicting OS using the five-gene signature. (F) A nomogram to estimate the annual survival rate of patients with adult soft-tissue sarcoma based on the five gene indicators. (G) Calibration plot for the nomogram-predicted probability of 5-year OS. (H) AUC of time-dependent ROC curves. ACTG2, actin γ2; APOD, apolipoprotein D; COPG2IT1, coatomer protein complex subunit γ2 imprinted transcript 1; COL6A6, collagen type VI α6 chain; OMD, osteomodulin; ROC, receiver operating characteristic; OS, overall survival; AUC, area under the curve.
Fig 5: IHC staining analysis of prognosis-related genes in the clinical validation cohort. (A) IHC staining of a high-risk case showing high expression of ACTG2 and low expression of APOD, COL6A6 and OMD, and a low-risk case displaying low expression of ACTG2 and high expression of APOD, COL6A6 and OMD (main image, ×40 magnification, and inset image, ×400 magnification). (B) KM curves of OS for ACTG2, APOD, COL6A6 and OMD based on the IHC staining results. (C) KM curves of PFS for ACTG2, APOD, COL6A6 and OMD. High expression of ACTG2 and low expression of APOD, COL6A6 or OMD were considered altered genes. (D) KM curves of OS for the number of altered genes. (E) KM curves of PFS for the number of altered genes. KM, Kaplan-Meier; IHC, immunohistochemistry; ACTG2, actin γ2; APOD, apolipoprotein D; COL6A6, collagen type VI α6 chain; OMD, osteomodulin; OS, overall survival; PFS, progression-free survival.
Supplier Page from Abcam for Anti-ACTG2 antibody