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Evaluation of TLS maturation and prognostic analysis of PSCC patients. A UMAP plot of the cell types identified in PSCC through spatial transcriptomic sequencing, depicting 7 distinct cell populations. B Dot plot displaying the expression of cell type-specific markers. The dot size represents the proportion of cells expressing the marker, while the colour intensity indicates the mean expression level. C Regions with AUCell scores for mTLS genes ( BCL6 , AICDA , CD38 , ICOS , CXCR5 , CXCL13 , and <t>CD21</t> ) above the median were defined as mTLSs, whereas those below the median were classified as iTLSs. D Spatial visualization of AUCell scores for mTLS genes, where warmer colours indicate higher scores. E Spatial distribution of iTLSs (green) and mTLSs (orange) across different regions of PSCC tissues. F Representative images H&E staining and immunofluorescence staining for CD20 (green) and CD21 (red) in iTLSs and mTLSs. G GO enrichment analysis of upregulated genes in mTLS. Darker colours denote more significant enrichment of the pathway, and larger dots represent a greater number of genes involved. H Kaplan–Meier survival curves showing that patients with mTLSs experienced significantly longer overall survival than those with iTLSs, as determined using ssGSEA ( P < 0.05)
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anti cd21 35 bv711 7e9 200 biolegend  (Revvity)
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Evaluation of TLS maturation and prognostic analysis of PSCC patients. A UMAP plot of the cell types identified in PSCC through spatial transcriptomic sequencing, depicting 7 distinct cell populations. B Dot plot displaying the expression of cell type-specific markers. The dot size represents the proportion of cells expressing the marker, while the colour intensity indicates the mean expression level. C Regions with AUCell scores for mTLS genes ( BCL6 , AICDA , CD38 , ICOS , CXCR5 , CXCL13 , and <t>CD21</t> ) above the median were defined as mTLSs, whereas those below the median were classified as iTLSs. D Spatial visualization of AUCell scores for mTLS genes, where warmer colours indicate higher scores. E Spatial distribution of iTLSs (green) and mTLSs (orange) across different regions of PSCC tissues. F Representative images H&E staining and immunofluorescence staining for CD20 (green) and CD21 (red) in iTLSs and mTLSs. G GO enrichment analysis of upregulated genes in mTLS. Darker colours denote more significant enrichment of the pathway, and larger dots represent a greater number of genes involved. H Kaplan–Meier survival curves showing that patients with mTLSs experienced significantly longer overall survival than those with iTLSs, as determined using ssGSEA ( P < 0.05)
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Evaluation of TLS maturation and prognostic analysis of PSCC patients. A UMAP plot of the cell types identified in PSCC through spatial transcriptomic sequencing, depicting 7 distinct cell populations. B Dot plot displaying the expression of cell type-specific markers. The dot size represents the proportion of cells expressing the marker, while the colour intensity indicates the mean expression level. C Regions with AUCell scores for mTLS genes ( BCL6 , AICDA , CD38 , ICOS , CXCR5 , CXCL13 , and <t>CD21</t> ) above the median were defined as mTLSs, whereas those below the median were classified as iTLSs. D Spatial visualization of AUCell scores for mTLS genes, where warmer colours indicate higher scores. E Spatial distribution of iTLSs (green) and mTLSs (orange) across different regions of PSCC tissues. F Representative images H&E staining and immunofluorescence staining for CD20 (green) and CD21 (red) in iTLSs and mTLSs. G GO enrichment analysis of upregulated genes in mTLS. Darker colours denote more significant enrichment of the pathway, and larger dots represent a greater number of genes involved. H Kaplan–Meier survival curves showing that patients with mTLSs experienced significantly longer overall survival than those with iTLSs, as determined using ssGSEA ( P < 0.05)
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Evaluation of TLS maturation and prognostic analysis of PSCC patients. A UMAP plot of the cell types identified in PSCC through spatial transcriptomic sequencing, depicting 7 distinct cell populations. B Dot plot displaying the expression of cell type-specific markers. The dot size represents the proportion of cells expressing the marker, while the colour intensity indicates the mean expression level. C Regions with AUCell scores for mTLS genes ( BCL6 , AICDA , CD38 , ICOS , CXCR5 , CXCL13 , and <t>CD21</t> ) above the median were defined as mTLSs, whereas those below the median were classified as iTLSs. D Spatial visualization of AUCell scores for mTLS genes, where warmer colours indicate higher scores. E Spatial distribution of iTLSs (green) and mTLSs (orange) across different regions of PSCC tissues. F Representative images H&E staining and immunofluorescence staining for CD20 (green) and CD21 (red) in iTLSs and mTLSs. G GO enrichment analysis of upregulated genes in mTLS. Darker colours denote more significant enrichment of the pathway, and larger dots represent a greater number of genes involved. H Kaplan–Meier survival curves showing that patients with mTLSs experienced significantly longer overall survival than those with iTLSs, as determined using ssGSEA ( P < 0.05)
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Evaluation of TLS maturation and prognostic analysis of PSCC patients. A UMAP plot of the cell types identified in PSCC through spatial transcriptomic sequencing, depicting 7 distinct cell populations. B Dot plot displaying the expression of cell type-specific markers. The dot size represents the proportion of cells expressing the marker, while the colour intensity indicates the mean expression level. C Regions with AUCell scores for mTLS genes ( BCL6 , AICDA , CD38 , ICOS , CXCR5 , CXCL13 , and CD21 ) above the median were defined as mTLSs, whereas those below the median were classified as iTLSs. D Spatial visualization of AUCell scores for mTLS genes, where warmer colours indicate higher scores. E Spatial distribution of iTLSs (green) and mTLSs (orange) across different regions of PSCC tissues. F Representative images H&E staining and immunofluorescence staining for CD20 (green) and CD21 (red) in iTLSs and mTLSs. G GO enrichment analysis of upregulated genes in mTLS. Darker colours denote more significant enrichment of the pathway, and larger dots represent a greater number of genes involved. H Kaplan–Meier survival curves showing that patients with mTLSs experienced significantly longer overall survival than those with iTLSs, as determined using ssGSEA ( P < 0.05)

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Association of CCL21 + CAFs with B-cell recruitment and TLS maturation in penile squamous cell carcinoma

doi: 10.1007/s00262-026-04348-5

Figure Lengend Snippet: Evaluation of TLS maturation and prognostic analysis of PSCC patients. A UMAP plot of the cell types identified in PSCC through spatial transcriptomic sequencing, depicting 7 distinct cell populations. B Dot plot displaying the expression of cell type-specific markers. The dot size represents the proportion of cells expressing the marker, while the colour intensity indicates the mean expression level. C Regions with AUCell scores for mTLS genes ( BCL6 , AICDA , CD38 , ICOS , CXCR5 , CXCL13 , and CD21 ) above the median were defined as mTLSs, whereas those below the median were classified as iTLSs. D Spatial visualization of AUCell scores for mTLS genes, where warmer colours indicate higher scores. E Spatial distribution of iTLSs (green) and mTLSs (orange) across different regions of PSCC tissues. F Representative images H&E staining and immunofluorescence staining for CD20 (green) and CD21 (red) in iTLSs and mTLSs. G GO enrichment analysis of upregulated genes in mTLS. Darker colours denote more significant enrichment of the pathway, and larger dots represent a greater number of genes involved. H Kaplan–Meier survival curves showing that patients with mTLSs experienced significantly longer overall survival than those with iTLSs, as determined using ssGSEA ( P < 0.05)

Article Snippet: The following antibodies were used: CD20 (Proteintech, 60,271–1-Ig, 1:500), CD21 (Proteintech, 28,206–1-AP, 1:250), ACTA2/α-SMA (Sigma, SAB5700835, 1:300), and CCL21 (Abcam, ab9851, 1:300).

Techniques: Sequencing, Expressing, Marker, Staining, Immunofluorescence