Stratification and pathway enrichment analysis of TCGA-BLCA based on ICD-related genes. A-C. Gene expression heatmap (A), clinical heatmap (B), and survival analysis (C) of ICD-high and ICD-low groups. D-E. Differential gene heatmap (D) and volcano plot (E) of ICD-high and ICD-low groups. F-G. GO enrichment analysis. H. KEGG enrichment analysis. I-L. GSEA enrichment analysis.

The immune characteristics within the tumor microenvironment vary between ICD-high and ICD-low groups. ESTIMATE scores (A), immune scores (B), stromal scores (C), and tumor cell purity (D) of ICD-high and ICD-low groups. E. Immune cell infiltration in the two subgroups. F-H. Expression profiles of the two subgroups in 13 immune-related pathways (F), 8 immune checkpoints (G), and HLA gene family (H). I. Efficacy of immunotherapy in the two subgroups.

Construction and validation of risk-scoring model. A. Hazard ratio of four ICD-related genes identified by Cox regression analysis. B-C. LASSO regression analysis for constructing the prognostic model. D-E. Survival analysis of high-risk and low-risk groups in TCGA (D) and GEO (E) databases. Analyzed the correlation between risk-scores and patients, along with four key genes, in both TCGA (F-H) and GEO databases (I-K).

The association between risk-scores and clinical features and immune characteristics. A-D. The association between the four genes composing the risk-scores and patient prognosis. E. Clinical heatmap of the two subgroups. F-G. Differences in stage (F) and grade (G) between the two subgroups. H-J. Constructed nomogram (H) and ROC curve (I-J). K-L. Cox regression analysis.M.Stromal scores, immune scores, and ESTIMATE scores of the high-risk and low-risk groups. N. Immune infiltration in the two subgroups.

The association between the risk-score and the sensitivity to chemotherapy (A-P) and immunotherapy (Q-T).

Validate the risk-scoring model through tissue arrays and single-cell sequencing. A-B. Analyzing the expression of CALR, IFN1B, IFNG, and IL1R1 using immunofluorescence staining. C-D. Survival analysis of the four genes (C) and risk-scores (D) in patients. E-I. The relationship between risk-scores and CD39, CD8+, CD8+LAG3+, Tumor Size, and T Stage. J-K. Expression analysis of CALR, IFN1B, IFNG, and IL1R1 in single-cell sequencing data.