Fig 1: Melanoma specimens reduce PRGs positive cells in epidermal clinically.A Overall survival analysis based on K-M was calculated to identify the potential PRGs clinically. B PPI via MCODE and CytoNCA algorithm was established, in which PRGs were screened out as central protein functioning in melanoma deficiency. C GZMA, GSDMB, CHMP4A, NLRP1, and IL18 protein expression of melanoma and control patients detected by ELISA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, n = 5, compared with the normal group. D Representative IHC stanning of GZMA+, GSMDB+, CHMP4A+, NLRP1+, and IL18+ in melanoma specimens from the clinic. Scale bar = 100 µm, n = 5 samples per group. E The quantification of PRGs protein in epidermal areas, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, compared with the control group.
Fig 2: GZMA+ cells and GSDMB+ cells are secreted by CD8+ T cells.A Representative multi-color staining of phenotypes of control and melanoma. 40,6-diamidino-2phenylindole (DAPI) (blue), CD8 (pink), GZMA (green), and GSDMB (red). Scale bars, 50 μm. B The scattergrams of different CD8+GZMA+and CD8 + GSDMB+ percent cells among the whole sample.
Fig 3: GZMA+ cells and GSDMB+ cells are secreted by NK T cells.A Representative multi-color staining of phenotypes of control and melanoma. 40,6-diamidino-2phenylindole (DAPI) DAPI (blue), CD57 (pink), GZMA (green), and GSDMB (red). Scale bars, 50 µm. B The scattergrams of different CD8+GZMA+, CD8+GSDMB+, CD57+GZMA+, and CD57+GSDMB+ percent cells among the whole sample.
Fig 4: PRGs were primarily expressed in immune cells.A Feature plots depicting the expression of key pyroptosis, violin plots were also displayed to determine the cell type. B Representative IHC stanning of GZMA+, GSMDB+, CHMP4A+, NLRP1+, and IL18+ in melanoma specimens, especially in lymphocyte areas. Scale bar = 100 µm, n = 4 samples per group. C The quantification of PRGs protein in dermal areas, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, compared with the control group.
Fig 5: Clinical relevance and individualized prognostic nomogram.A Forest plot of univariate Cox regression of the PRGs, including pyroptosis (GZMA and GSDMB) and clinical predicts (age, gender, pT/N/M-stage). B Forest plot of multivariate Cox regression of the PRGs, including pyroptosis (GZMA and GSDMB) and clinical predicts (age, gender, pT/N/M-stage). C K–M analysis showing the overall survival rate of high-risk patients (red) and low-risk patients (blue). The numbers of patients and the risk classification are indicated in the figure. D ROC curve was adopted to evolute the prediction performance of pyroptosis-related signature. E The individual prognostic nomogram of the 1-year, 2-year, and 3-year survival prediction of melanoma patients. F Calibration curve for the individual prognostic nomogram. A dashed diagonal line represents the ideal nomogram, and the blue line, red line, and orange line represent the 1-y, 2-y, and 3-y observed nomograms.
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