mihc staining  (Boster Bio)

Journal: Signal Transduction and Targeted Therapy

Article Title: Prostaglandin E 2 -driven dedifferentiation of Schwann cells leads to perineural invasion in pancreatic ductal adenocarcinoma

doi: 10.1038/s41392-026-02648-x

Figure Lengend Snippet: SCs are enriched in the PNI-positive region. a Schematic diagram of the workflow; some of the graphical elements were created wtith BioGDP. b Division of perineural invasion regions (PNI-region: PNI, PNI-near, PNI-mid, PNI-away) and nonperineural invasion regions (NPNI: NPNI, NPNI-near, NPNI-mid, NPNI-away) in representative spatial transcriptomic images of PDAC PNI samples, with corresponding representative HE-stained images of spot points (right) (n = 4). N: Nerve, T: Tumor. Scale bar: 500 μm. c GSVA of the PNI region and the NPNI region. d Correlations between different cell types and the expression levels of marker genes. e Cell type distribution scatter plot based on the UMAP dimensionality reduction algorithm; f UMAP plot of SC subsets in pancreatic cancer tissues. g Feature score of nomyelinating SCs at the single-cell level in each of the two SC subsets ( p = 0.025). h Representative images of RCTD single-cell mapping results based on PNI regions and NPNI regions (n = 4); i Stacked bar charts of different PNI regions and NPNI regions, showing the proportions of various cell types in different samples. j , k Representative mIHC images of human normal pancreatic tissues, PDAC-PNI- tissues, and PDAC-PNI+ tissues stained with PGP9.5, SOX2, c-Jun, and MUC1 antibodies. Scale bar: 100 μm (left), 50 μm (right). The ratios of SOX2+ and c-Jun+ positive cells in normal pancreatic tissues, PDAC-PNI- tissues, and PDAC-PNI+ tissues ( n = 5). All the results are presented as the means ± SDs. Each data point in the bar graphs represents an individual sample. Statistical significance was determined by a two-tailed unpaired t test (Fig. 1g) and one-way analysis of variance followed by Tukey’s HSD post hoc test (Fig. 1j). **** p < 0.0001

Article Snippet: For mIHC staining, formalin-fixed sections were stained with antibodies against PGP9.5 (1:200, Abways, CY6722), MUC1 (1:200, BOSTER, BM4548), SOX2 (1:200, Diagbio, db16528), c-Jun (1:200, Selleck, F0168), and p75NTR (1:200, ABclonal, A11169) according to the manufacturer’s instructions for the multicolor immunofluorescence kit (Zolgene, PR-004507) and imaged with a KFbio Pathology Imaging System (KF-FL-005).

Techniques: Staining, Expressing, Marker, Single Cell, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Prostaglandin E 2 -driven dedifferentiation of Schwann cells leads to perineural invasion in pancreatic ductal adenocarcinoma

doi: 10.1038/s41392-026-02648-x

Figure Lengend Snippet: Elevation of PTGES/PGE 2 in the coculture system is correlated with PNI. a Venn diagram showing the intersection of multiomics data identifying PTGES as a key gene in PNI. b Expression of PTGES genes via spatial transcriptome analysis. Scale bar: 500 μm ( n = 4). c Violin plot showing the expression of PTGES in various PNI regions and NPNI regions via spatial transcriptomics ( n = 4). d Representative mIHC images of human normal pancreatic tissues, PDAC-PNI- tissues, and PDAC-PNI+ tissues stained with PGP9.5, p75NTR and MUC1 antibodies ( n = 3). Scale bar: 100 μm (left), 50 μm (right). e IHC staining of PTGES in human normal pancreatic tissues, PDAC-PNI- tissues, and PDAC-PNI+ tissues ( n = 9), N: nerve, T: tumor. Scale bar: 100 μm. l H-scores of PTGES. f , i Western blotting was performed to detect the protein expression level of PTGES in human normal pancreatic tissues and PDAC tissues ( n = 3), N: normal tissue, T: tumor tissue. g , j Western blotting was used to determine the protein expression level of PTGES in PANC-1 and BxPC-3 cells after coculture with RSC96 cells ( n = 3). h , k Western blotting was conducted to measure the protein expression level of PTGES in PANC-1 and BxPC-3 cells after coculture with sNF96.2 cells ( n = 3). m ELISA was used to detect the concentration of PGE 2 in the culture medium ( n = 3), and some of the graphical elements were created wtith Figdraw. All the results are presented as the means ± SDs. Each data point in the graphs represents an individual sample. Statistical significance was determined via two-tailed unpaired t tests (for Fig. 4i–k), one-way analysis of variance followed by Tukey’s HSD post hoc test (for Fig. 4m) and the Kruskal‒Wallis H test followed by Dunn’s multiple comparisons test (for Fig. 4l). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: For mIHC staining, formalin-fixed sections were stained with antibodies against PGP9.5 (1:200, Abways, CY6722), MUC1 (1:200, BOSTER, BM4548), SOX2 (1:200, Diagbio, db16528), c-Jun (1:200, Selleck, F0168), and p75NTR (1:200, ABclonal, A11169) according to the manufacturer’s instructions for the multicolor immunofluorescence kit (Zolgene, PR-004507) and imaged with a KFbio Pathology Imaging System (KF-FL-005).

Techniques: Expressing, Spatial Transcriptomics, Staining, Immunohistochemistry, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay, Two Tailed Test

Journal: Clinical & Translational Immunology

Article Title: Plasma EBV quantification is associated with the efficacy of immune checkpoint blockade and disease monitoring in patients with primary pulmonary lymphoepithelioma‐like carcinoma

doi: 10.1002/cti2.1515

Figure Lengend Snippet: Correlation between PD‐L1 expression and TMB and plasma EBV DNA before ICB treatment in PLELC. (a) Proportion of different PD‐L1 expression in the EBNA‐1 high group ( EBNA‐1 > 11 231 copies mL −1 before immunotherapy) vs. the EBNA‐1 low group ( EBNA‐1 ≤ 11 231 copies mL −1 ). (b) PFS of patients with ICB‐treated PLELC with both high plasma EBNA‐1 before immunotherapy and PD‐L1 ≥ 1% vs. others. (c) PFS of patients with ICB‐treated PLELC with both high plasma BamHI‐W before immunotherapy and PD‐L1 ≥ 1% vs. others. (d) PFS of 23 patients with ICB‐treated PLELC with TMB < 10/MB vs. ≥10/MB. (e) Scatter plot of TMB and lg ( EBNA‐1 ). (f) Scatter plot of TMB and lg ( BamHI‐W ). (g) Median TMB of the EBNA‐1 high group ( EBNA‐1 > 11 231 copies mL −1 before immunotherapy) vs. the EBNA‐1 low group ( EBNA‐1 ≤ 11 231 copies mL −1 ). (h) Median TMB of the BamHI‐W high group ( BamHI‐W ≥ 10 446 copies mL −1 before immunotherapy) vs. the BamHI‐W low group ( BamHI‐W < 10 446 copies mL −1 ). (i) Representative mIHC staining for PD‐1, PD‐L1, LAG3, TIM3 and CD3 (panel 1; left), and CD4, CD8, CD68, FOXP3 and CK (panel 2; right). (j) Heatmap of correlation analysis of lg (EBNA‐1) and CD3, CD4, CD8, PD1, PD‐L1, TIM3, LAG3, FOXP3, CK5‐6, CD68. (k) Scatter plot of positive percentage of lg ( EBNA‐1 ) and CD8 in 15 patients with advanced PLELC.

Article Snippet: Tissue samples from 15 patients with advanced PLELC were used to detect the lymphocytes markers CD3, LAG3, PD1, PD‐L1, TIM3, CD4, CD8, CD68, CK and FOXP3 by mIHC staining (Genecast Biotechnology Co., Ltd, Wuxi, China).

Techniques: Expressing, Staining