Fig 1: SERPINE2 serves as a metastasis-associated oncogene in RCC and drug response.A The Oncoprint of conventional marker genes of RCC with alterations in SERPINE2 high and low groups. Tumor mutation burden is represented for individual samples as a bar chart above the oncoprint. B The GSEA plot shows enrichment of EMT-related pathways in SERPINE2 high group in TCGA. FDR < 0.05 is considered as significantly enriched. C SERPINE2 expression between tumor tissues from patients with metastasis (positive N stage or M stage >0, n = 89) and from patients with no metastasis (N stage or M stage = 0, n = 199). Data seen in Table S10. D SERPINE2 expression among primary (n = 9), metastatic tumors (n = 26) and normal tissues (n = 9) in GSE105261. Data were obtained from https://www.aclbi.com/static/index.html#/geo. E SERPINE2 expression between tumors (n = 72) and normal tissues (n = 72) in GSE53757. Data were obtained from https://www.aclbi.com/static/index.html#/geo. F IC50 prediction of sunitinib for SERPINE2 high and low groups in TCGA, based on the GDSC database. G The estimate score of tumor tissues from SERPINE2 high and low groups, reflecting the immune infiltration. H MTOR expression from cells in scRNA-seq analysis (50%: 50%). I UMAP plot shows the terminally exhausted CD8+ T cell signature expression. J The KM survival curve of SERPINE2-high and SERPINE2-low groups in the cohort of clinical trials Checkmate025 with Nivolumab. *p < 0.05; **p < 0.01; ***p < 0.001.
Fig 2: SERPINE2 predicts poor survival and RCC metastasis in our clinical cohort.A Representative IHC staining and H-score for SERPINE2 in RCC tissues and matched para-tumor tissues (scale bar = 50 µm). B IHC score between tumor tissues and matched para-tumor tissues. C IHC score between tumor tissues from patients with metastasis and no metastasis. D IHC score between patients with stage I, II and stage III, IV. E IHC score between patients with Fuhrman I, II and Fuhrman III, IV. F The ROC curve for SERPINE2 expression in 5-year OS (TMA30 cohort). G The KM curve for OS difference between SERPINE2 high and low groups. H The KM curve for PFS difference between SERPINE2 high and low groups. I The nomogram for 1-, 3- and 5-year OS. OS overall survival, PFS progression-free survival.
Fig 3: SERPINE2 promotes ccRCC progression, accompanied by EMT.A, B Representative images of transwell assay without or with Matrigel shows the migration and invasion abilities among shCtrl, shSERPINE2#1 and shSERPINE2#2 group in Caki-1 cells, relative change compared with shCtrl group. Scale bar = 100 µm. C, D Representative images of transwell assay without or with Matrigel shows the migration and invasion abilities between Vector, OE-SERPINE2 group in 786-O cell, relative change compared with Vector group. Scale bar = 100 µm. E The cell survival rate among shCtrl, shSERPINE2#1 and shSERPINE2#2 group in Caki-1 cells, relative change compared with shCtrl group. F The cell survival rate among Vector, OE-SERPINE2 group in 786-O cell, relative change compared with Vector group. G Whole body bioluminescence (photons/second) following tail vein injection of cells in mice. Differences occurred from day 7 time point (Green line). H The lung tissue of 786-O-OE-SERPINE2 or 786-O-NC-SERPINE2 of mice; the representative lung metastasis site by HE staining. Scale bar = 500 µm; Scale bar = 50 µm. The IHC results of CA9 and SERPINE2; Scale bar = 50 µm; Scale bar = 20 µm. EMT epithelial-mesenchymal transition.
Fig 4: NNMF clustering characterizing heterogenic malignant cells and identifying metastasis program in RCC tumors.A The heatmap presents DEGs (rows) identified by NMF, which are clustered by expression in each cell (column) of the representative Malignant 4. The gene signature was identified and indicated on the top. B The heatmap shows the correlations of 60 intratumoral procedures from six malignant subsets. The clustering identified six coherent expression programs across tumors. C The heatmap depicts genes scores by each sample (column) for genes (row) in the Metastasis-II program. D Venn plot shows the number of DEGs (malignant vs. non-malignant cells in scRNA profile) and the top genes in the NNMF program. E The KM survival curve of SERPINE2-high and SERPINE2-low group in TCGC KIRC (Left, n = 530); the SERPINE2 expression between tumor tissues (n = 530) and normal tissues (n = 72) in KIRC (Right). *p < 0.05; **p < 0.01; ***p < 0.001. F Violin plot shows the SERPINE2 expression among Malignant 1–6 subsets. G Violin plot shows the SERPINE2 expression among cells from primary and metastatic tumors and non-malignant subsets. H The comprehensive plot of Caki-1 and HK-2 cells shows the compartment status at the SERPINE2 locus B to A, and this locus at ATAC-seq presents an open status. The red bar represents A compartment; Blue bar represents B compartment. Merge bar represents the compartment status alternations. The Diff_BPM shows all the DEGs between the two cell lines in this chromatin region. I The TAD analysis shows TAD boundaries reforming/disappearing and insulation scores between HK-2 and Caki-1. Red line, for Caki-1; Green line, for HK2. Caki-1 shows lower insulation score than HK-2 do, and new boundary reforms compared with HK-2. DEGs differentially expressed genes.
Supplier Page from Abcam for Anti-SERPINE2/PN-1 antibody [EPR8776]