Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A The expression of neutrophils and CD8 + T cells in low- and high-glycolysis clusters, divided by ssGSEA score of glycolysis-related genes, in TCGA and CPTAC PAAD database. Data are presented as box plots showing the median (center line), the first and third quartiles (box bounds), and the whiskers extend to 1.5 times the interquartile range from the box. B The correlation of glycolysis level and neutrophils or CD8 + T cells expression in TCGA ( n = 179 patients) and CPTAC ( n = 140 patients) PAAD database. C Kaplan–Meier plots representing survival probabilities in TCGA-PAAD patients according to the relative level of glycolysis-related or neutrophil-related gene expression. D The diagram illustrating the targets of 2DG and shRNA in the glycolysis process. E CyTOF analysis of tumor-infiltrating immunocytes in subcutaneous tumor models with or without 2DG treatment: tSNE plots showing 12 meta-clusters based on the expression of 41 markers for the immunocytes. F Bar chart of the frequencies of the immune cell subsets in two experimental groups. G A schematic diagram showing the orthotopic PDAC model ( n = 6 mice per group in one experiment) with or without 2DG treatment. H , I Image and weights of the orthotopic tumors in the experimental groups from ( G ) at the end of the experiments. ( n = 6 mice per group). ( J – M ) Tumor-infiltrating neutrophils, CD8 + T cells, GZMB + CD8 + T cells, and PD-1 + CD8 + T cells isolated from the orthotopic tumors were analyzed using flow cytometry: representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 6 mice per group). N A schematic diagram showing the orthotopic tumor model in C57BL/6 J mice ( n = 6 mice per group in one experiment) treated with or without anti-Ly6G/anti-CD8 antibody. O , P Image and weights of the orthotopic tumors in the experimental groups from ( N ) at the end of the experiments ( n = 6 mice per group). Q – S Tumor-infiltrating neutrophils, CD8 + T cells, and GZMB + CD8 + T cells isolated from the orthotopic tumors were analyzed using flow cytometry: Representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 6 mice per group). T , U Representative luminescence images of the mouse model in ( N ) and statistical analysis of the total flux ( n = 6 mice per group). Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( A, I–M, P–S, and U ), two-tailed Pearson’s correlation analysis ( B ), and log-rank test ( C ). Source data are provided as a Source Data file.

Article Snippet: The antibodies used here included anti-L-Lactyl-Histone H3K18 antibody (1:500, PTMBIO, #PTM-1406RM), anti-GRO antibody (1:200, AFFINITY, #AF5403), Ki-67 (1:400, Servicebio, # GB111499 ), anti-Ly6G antibody (1:500, Servicebio, #GB11229), and anti-CD8 antibody (1:200, Cell Signaling Technology, #70306), and anti-PCAF antibody(1:200, Cell Signaling Technology, #3378).

Techniques: Expressing, Gene Expression, shRNA, Isolation, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A The flowchart illustrates that RNA sequencing was conducted using two PDAC cell lines with or without 2DG treatment. Subsequently, the intersection of detected genes within the chemokine family in both cell lines is identified, and a heatmap is generated to display the log 2 fold change (2DG versus vehicle) of these genes. B , C Relative mRNA levels of Cxcl1 in PDAC cell lines following glycolysis inhibition, either by using 2DG or silencing LDH, were analyzed by qRT-PCR ( n = 3 independent experiments). D , E Relative protein levels of CXCL1 in PDAC cell lines following glycolysis inhibition, either by using 2DG or silencing LDH, were determined using ELISA assay ( n = 3 independent experiments). F Representative IHC images of subcutaneous tumors ( n = 6 mice for 2DG treatment, n = 5 mice for shLDH/shNTC groups) stained by CXCL1 antibody (scale bars = 100 μm). G Relative serum CXCL1 levels in mice treated with or without 2DG were measured by ELISA ( n = 6 mice per group). H Relative serum CXCL1 levels in healthy donors and PDAC patients were measured by ELISA (22 healthy samples and 27 PDAC samples). I Schematic diagram showing the in vitro migration assay: human/mouse neutrophils were co-incubated with the culture medium supernatant from PDAC cells treated with 2DG or LDH-knockdown. J Neutrophils were co-cultured with CD8 + T cells in different proportions, and the proliferation of CD8 + T cells was detected with the CFSE assay ( n = 6 biologically independent samples). K The migratory activity of neutrophils co-incubated with the culture medium supernatant from PDAC cells treated with shLDH and recombinant CXCL1 was analyzed by counting the penetrated cell numbers ( n = 3 biologically independent samples). L The neutrophils were treated with SX-682 or Navarixin for 1.5 h in advance, and the migratory activity of neutrophils co-incubated with the culture medium supernatant from PDAC cell lines treated with shLDH was analyzed by counting the penetrated cell numbers ( n = 3 biologically independent samples). M Representative luminescence images of the orthotopic tumor in mice and statistical analysis of MFI ( n = 5 mice per group). N Tumor-infiltrating neutrophils isolated from the orthotopic tumors were analyzed using flow cytometry ( n = 5 mice per group). Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test. Source data are provided as a Source Data file.

Article Snippet: The antibodies used here included anti-L-Lactyl-Histone H3K18 antibody (1:500, PTMBIO, #PTM-1406RM), anti-GRO antibody (1:200, AFFINITY, #AF5403), Ki-67 (1:400, Servicebio, # GB111499 ), anti-Ly6G antibody (1:500, Servicebio, #GB11229), and anti-CD8 antibody (1:200, Cell Signaling Technology, #70306), and anti-PCAF antibody(1:200, Cell Signaling Technology, #3378).

Techniques: RNA Sequencing, Generated, Inhibition, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, In Vitro, Migration, Incubation, Knockdown, Cell Culture, CFSE Assay, Activity Assay, Recombinant, Isolation, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A Molecular docking simulated the binding affinities of PCAF for acetyl-CoA and lactyl-CoA. B Co-IP was performed to confirm the interaction between H3K18la and PCAF in PANC1 and KPC cells. PCAF and H3K18la were detected on the same gel. C, D Western blot analysis demonstrates the level of H3K18la in PDAC cells with or without the treatment of PCAF inhibitors (bromosporine or Embelin). Histone H3 and H3K18la blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. E Western blot analysis demonstrates the level of H3K18la in PDAC cells following the knockdown of Pcaf . Histone H3, PCAF, β-actin and H3K18la blots are from parallel-processed separate gels with samples from the same experiment. F Western blots of in vitro histone acetylation or lactylation assay, the samples as indicated. Blots are from parallel-processed separate gels with samples from the same experiment. G , H Relative Cxcl1 RNA and protein levels with the vehicle or bromosporine treatment were analyzed by qRT-PCR and ELISA ( n = 3 independent experiments), respectively. I Relative Cxcl1 RNA levels after Pcaf knockdown were analyzed by qRT-PCR ( n = 3 independent experiments). J A schematic diagram showing the orthotopic tumor construction in C57BL/6 J mice ( n = 5 mice per group in one experiment, 2 × 10 6 KPC-luc cells per mouse) with or without bromosporine treatment. K , L Image and weights of the orthotopic tumors in the experimental groups from ( J ) at the end of the experiments ( n = 5 mice per group). M Lactylation levels of H3K18 and PanK in orthotopic tumors with or without bromosporine treatment were detected by western blot ( n = 4 biologically independent samples per group). Histone H3, H3K18la, and PanKla blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. N Serum CXCL1 levels in mice with or without bromosporine treatment were measured by ELISA ( n = 5 mice per group). O – R Tumor-infiltrating neutrophils, CD8 + T cells, GZMB + CD8 + T cells, and PD-1 + CD8 + T cells isolated from the orthotopic tumors were analyzed using flow cytometry. Representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 5 mice per group). S The migratory abilities of neutrophils co-incubated with the culture medium supernatant from shNTC/ Pcaf PDAC cells treated with recombinant CXCL1 were analyzed by counting the penetrated cell numbers ( n = 3 biologically independent samples). T Representative IHC images stained by CXCL1 or H3K18la antibody (scale bar = 100 μm, left panel), correlation analysis of H-scores of CXCL1 and H3K18la in PDAC samples ( n = 21 patients, right panel). Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( G – I , L , and N – S ) and two-tailed Pearson’s correlation analysis ( T ). Unless otherwise indicated, all western blots had three independent experimental repetitions with consistent results. Source data are provided as a Source Data file.

Article Snippet: The antibodies used here included anti-L-Lactyl-Histone H3K18 antibody (1:500, PTMBIO, #PTM-1406RM), anti-GRO antibody (1:200, AFFINITY, #AF5403), Ki-67 (1:400, Servicebio, # GB111499 ), anti-Ly6G antibody (1:500, Servicebio, #GB11229), and anti-CD8 antibody (1:200, Cell Signaling Technology, #70306), and anti-PCAF antibody(1:200, Cell Signaling Technology, #3378).

Techniques: Binding Assay, Co-Immunoprecipitation Assay, Western Blot, Knockdown, In Vitro, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Isolation, Flow Cytometry, Incubation, Recombinant, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A A schematic diagram showing the subcutaneous tumor model ( n = 6 mice per group in one experiment, 6 × 10 6 PANC02 cells per mouse) treated with bromosporine and anti-PD-1 antibody. B. The tumor volume growth curves of subcutaneous tumors from ( A ). C , D Image and weights of the subcutaneous tumors at the end point of experiments ( n = 6 mice per group). E The statistical analysis of the cell ratio of tumor-infiltrating neutrophils and CD8 + T cells isolated from subcutaneous tumors ( n = 3 mice per group). F Western blot analysis demonstrates H3K18la levels in the subcutaneous tumors ( n = 3 biologically independent samples per group) from ( A ). Histone H3 and H3K18la blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. G Relative serum CXCL1 levels in mice treated with or without bromosporine/anti-PD-1 antibody were measured by ELISA ( n = 6 mice per group). H A schematic diagram showing the combinational treatment schedule for the orthotopic KPC-luc tumor model ( n = 21 mice per group in one experiment, 2 × 10 6 KPC-luc cells per mouse). I , J Image and weights of the orthotopic tumors at the end point of experiments ( n = 6 mice per group). K – N Tumor-infiltrating neutrophils, CD8 + T cells, GZMB + CD8 + T cells, and PD-1 + CD8 + T cells were analyzed using flow cytometry. Representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 6 mice per group). O Western blot analysis showing the H3K18la levels in the orthotopic tumors ( n = 3 biologically independent samples per group) from ( H ). Histone H3 and H3K18la blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. P Representative luminescence images and Quantification of radiance intensity of the mouse model in ( H ). Q Survival probability of mice with orthotopically transplanted PDAC ( n = 15 mice per group). R A working model displaying the signaling pathway through which the aerobic glycolysis-mediated Lactate-PCAF-H3K18la-CXCL1 axis modulates the tumor microenvironment in pancreatic cancer, and the scientific basis for the development of a novel therapeutic strategy for PDAC. Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( B , D , E , G , J – N , P ) and the log-rank test ( Q ). Source data are provided as a Source Data file.

Article Snippet: The antibodies used here included anti-L-Lactyl-Histone H3K18 antibody (1:500, PTMBIO, #PTM-1406RM), anti-GRO antibody (1:200, AFFINITY, #AF5403), Ki-67 (1:400, Servicebio, # GB111499 ), anti-Ly6G antibody (1:500, Servicebio, #GB11229), and anti-CD8 antibody (1:200, Cell Signaling Technology, #70306), and anti-PCAF antibody(1:200, Cell Signaling Technology, #3378).

Techniques: Isolation, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: A spatially resolved atlas of gastric cancer characterises a lymphocyte-aggregated region

doi: 10.1038/s41467-026-68612-z

Figure Lengend Snippet: a Dot plot shows the intensity of T cell activation-associated signalling pathways across ST regions. b Schema for spatial blocks. c Comparisons of signalling pathways principal in T cell activation between spatial regions using normalised enrichment score. The top violin plots compare the signalling intensity between spatial regions, while the bottom bar plots indicate the percentage of spots enriched for the above signalling pathways for each spatial region ( P < 0.05, two-sided Wilcoxon test). Centre lines denote median values; whiskers denote 1.5 × the interquartile range; the lower and upper hinges represent the 25th and 75th percentiles, respectively. A total of 4,321 spatial blocks across 33 samples were used for comparison. d Lollipop plot indicates the Pearson coefficient measured correlations between the cellular abundance of T cell subsets and two T cell activation-associated signalling cascades within the LAR region. The Benjamini–Hochberg adjusted P value (two-sided) shows the statistical significance of each association. e The Pearson coefficient measured associations between the cellular abundance of CD4-Tem-GZMK and CD8-Tn-CCR7 with signals of TCR and Proliferation signalling pathways in the LAR region. Pearson correlation coefficients and two-sided P values are shown. The marginal rug indicates the 95% confidence interval. NES, normalised enrichment score. f Immunohistochemistry staining of CD8A, TCF1, and KI67 for the sample D16T2. Left panel, the whole slide view of H&E and mIHC images; middle panel, zoom-in views for an LAR highlighted by a rectangle in the left panel (scale bar, 100 μm); right panel, zoom-in views for the rectangle areas in the middle panel (scale bar, 10 μm). Triangles indicate TCF1 + KI67 + CD8A + cells . TCF1 + KI67 + CD8A + cells were observed within LARs of 11 samples in the main study cohort and five samples from an independent validation cohort.

Article Snippet: The protein markers used for staining were CD8 (1:400, clone C8/144B, 70306, Cell Signalling Technology), TCF1 (1:200, clone C63D9, 14456, Cell Signalling Technology), Ki67 (1:200, clone SP6, RM-9106-s1, Thermo Scientific), CD27 (1:2000, clone LPFS2/1611, ab268144, Abcam), CD70 (1:400, clone E3Q1A, 69209, Cell Signalling Technology), PD-1 (1:300, clone 29 F.1A12, 135210, Biolegend) and LAMP3 (1:200, clone 1010E1.01, Novus Biologicals).

Techniques: Activation Assay, Comparison, Immunohistochemistry, Staining, Biomarker Discovery

Journal: Nature Communications

Article Title: A spatially resolved atlas of gastric cancer characterises a lymphocyte-aggregated region

doi: 10.1038/s41467-026-68612-z

Figure Lengend Snippet: a Distinct immune checkpoint ligand-receptor pairs enriched in different spatial regions. * indicates P < 0.001. b Forest plot delineates the association between the axis of CD27-CD70 and donor groups based on effect sizes and their 95% confidence intervals. c Comparison of expression of CD27-CD70 between LAR blocks of Groups A and B ( P < 0.001, two-sided t-test). A total of 520 LAR spatial blocks across 26 samples were used for comparison. For all the box plots in this figure, the centre line of the box denotes the median value, and the horizontal line denotes the 95% confidence interval. d Boxplots compare the expression of inhibitory axes between NCR blocks of Groups A and B ( P < 0.001, two-sided t-test). A total of 1204 NCR spatial blocks across 33 samples were used for comparison. e Dot plot indicates the cellular communications of cDC-LAMP3 with T cell subsets coordinated by the axis of CD27-CD70 inferred from the scRNA-seq data. Benjamini–Hochberg adjusted P values were generated from a one-sided Z-test. f Immunohistochemistry staining of CD8A, PD1, CD27, LAMP3 and CD70 for the sample D11T1. The physical contact between these two cell types was observed within LARs of 11 samples from the main study cohort and five samples from the independent validation cohort. Left panel, the whole slide view of H&E and mIHC images; middle panel, zoom-in views for an LAR region highlighted by the rectangle of the left panel (scale bar, 100 μm); right panel, zoom-in views for the rectangle areas in the middle panel (scale bar, 10 μm). Yellow triangles indicate PD1 + CD27 + CD8 + cells, while white triangles indicate LAMP3 + CD70 + cells. g A Venn diagram illustrates the LAR spatial blocks expressing both axes of CD28 and CTLA4. h , Box plot compares the average distance between PD1 + CD27 + CD8 + and CD70 + LAMP3 + cells and the average distance between DAPI and CD70 + LAMP3 + cells within each LAR in the main study cohort. A total of 19 LAR regions across 10 samples were used for comparison. The Bonferroni-adjusted P value indicates the statistical significance between these two groups ( P < 0.001, two-sided t-test).

Article Snippet: The protein markers used for staining were CD8 (1:400, clone C8/144B, 70306, Cell Signalling Technology), TCF1 (1:200, clone C63D9, 14456, Cell Signalling Technology), Ki67 (1:200, clone SP6, RM-9106-s1, Thermo Scientific), CD27 (1:2000, clone LPFS2/1611, ab268144, Abcam), CD70 (1:400, clone E3Q1A, 69209, Cell Signalling Technology), PD-1 (1:300, clone 29 F.1A12, 135210, Biolegend) and LAMP3 (1:200, clone 1010E1.01, Novus Biologicals).

Techniques: Comparison, Expressing, Generated, Immunohistochemistry, Staining, Biomarker Discovery