lactate dehydrogenase b  (Proteintech)

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 Lactylation levels of pan-lysine and different lysine sites in histone H3 of 7 pairs of PDAC tissue and adjacent normal tissue were detected by western blot. B Representative IHC images of clinical PDAC and adjacent normal tissue stained by H3K18la antibody ( n = 79 patients). C The diagram illustrating the targets of 2DG, shRNA, and rotenone in the glycolysis process. D , E Relative levels of H3K18la and PanKla in PDAC cells treated with gradient doses of 2DG ( D ) or rotenone ( E ) for 24 h were detected by western blot. F Relative levels of H3K18la and PanKla in subcutaneous tumors treated with or without 2DG were detected by western blot. ( n = 4 biologically independent samples per group). G H3K18la levels of subcutaneous tumors treated with or without 2DG were detected by immunofluorescence staining. H The H3K18la levels in PDAC cells treated with 2DG or sodium lactate were detected by western blot. I The H3K18la level in PANC1 and PANC02 cells following LDH-knockdown and sodium lactate treatment by western blot. J Schematic diagram of the CUT&Tag assay for PanKla or H3K18la in BxPC3 cells with or without 2DG treatment. K TSS heatmaps presenting the binding density of PanKla and H3K18la with or without 2DG treatment. L Venn diagram indicating the overlap of differentially expressed genes in TCGA-PAAD between PDAC and adjacent normal tissue, differentially expressed genes in RNA-seq after 2DG treatment, and differentially bound genes of H3K18la and PanKla after 2DG treatment. M Genome browser track analysis shows the PanKla and H3K18la levels in the Cxcl1 promoter region with or without 2DG treatments. N The Cxcl1 promoters that are bound by H3K18la and PanKla in PDAC cells were quantified using ChIP-qPCR assays ( n = 3 independent experiments). O Relative mRNA levels of Cxcl1 in PDAC cells treated with 2DG or sodium lactate were detected by qRT-PCR ( n = 3 independent experiments). P Representative IHC images stained by CXCL1 or H3K18la antibody and correlation analysis of H-scores of CXCL1 and H3K18la in PDAC samples ( n = 21 patients). Scale bar = 100 μm. Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( N–O ) and two-tailed Pearson’s correlation analysis ( P ). Unless otherwise indicated, all western blots had three independent experimental repetitions with consistent results, where Histone H3, H3K9la, H3K14la, H3K18la, and PanKla blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. Source data are provided as a Source Data file.

Article Snippet: The primary antibodies used here included anti-LDHA antibody (Proteintech, #19987-1-AP, 1:3000), anti-LDHB antibody (Proteintech, #14824-1-AP, 1:5000), anti-H3 antibody (Proteintech, #17168-1-Ap, 1:3000), anti-PCAF antibody (Cell Signaling Technology, #3378S, 1:1000), anti-P300 antibody (Proteintech, #20695-1-AP, 1:500), anti-GCN5 antibody (Proteintech, #66575-1-Ig, 1:3000), anti-beta Actin antibody (PTMBIO, #PTM-5028, 1:1000), anti-human CXCL1 Polyclonal antibody (Proteintech, #12335-1-AP), anti-mouse CXCL1 Polyclonal antibody (Proteintech, #30322-1-AP), anti-L-Lactyl Lysine antibody (PTMBIO, #PTM-1401RM, 1:1000), anti-L-Lactyl-Histone H3K18 antibody (PTMBIO, #PTM-1406RM, 1:1000), anti-L-Lactyl-Histone H3K9 antibody (PTMBIO, #PTM-1419RM, 1:1000), anti-L-Lactyl-Histone H3K14 antibody (PTMBIO, #PTM-1414RM, 1:1000), anti-H4 antibody (PTMBIO, #PTM-1009, 1:1000), anti-L-Lactyl-Histone H4K12 antibody (PTMBIO, #PTM-1411RM, 1:1000), anti-L-Lactyl-Histone H4K8 antibody (PTMBIO, #PTM-1415RM, 1:1000), anti-Acetyl-Histone H3 (Lys18) antibody (PTMBIO, #PTM-114RM, 1:1000), anti-Acetyllysine antibody (PTMBIO, #PTM-101, 1:1000), and anti-Acetyl-Histone H4 (Lys8) antibody (PTMBIO, #PTM-190, 1:1000).

Techniques: Western Blot, Staining, shRNA, Immunofluorescence, Knockdown, Binding Assay, RNA Sequencing, ChIP-qPCR, Quantitative RT-PCR, 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 primary antibodies used here included anti-LDHA antibody (Proteintech, #19987-1-AP, 1:3000), anti-LDHB antibody (Proteintech, #14824-1-AP, 1:5000), anti-H3 antibody (Proteintech, #17168-1-Ap, 1:3000), anti-PCAF antibody (Cell Signaling Technology, #3378S, 1:1000), anti-P300 antibody (Proteintech, #20695-1-AP, 1:500), anti-GCN5 antibody (Proteintech, #66575-1-Ig, 1:3000), anti-beta Actin antibody (PTMBIO, #PTM-5028, 1:1000), anti-human CXCL1 Polyclonal antibody (Proteintech, #12335-1-AP), anti-mouse CXCL1 Polyclonal antibody (Proteintech, #30322-1-AP), anti-L-Lactyl Lysine antibody (PTMBIO, #PTM-1401RM, 1:1000), anti-L-Lactyl-Histone H3K18 antibody (PTMBIO, #PTM-1406RM, 1:1000), anti-L-Lactyl-Histone H3K9 antibody (PTMBIO, #PTM-1419RM, 1:1000), anti-L-Lactyl-Histone H3K14 antibody (PTMBIO, #PTM-1414RM, 1:1000), anti-H4 antibody (PTMBIO, #PTM-1009, 1:1000), anti-L-Lactyl-Histone H4K12 antibody (PTMBIO, #PTM-1411RM, 1:1000), anti-L-Lactyl-Histone H4K8 antibody (PTMBIO, #PTM-1415RM, 1:1000), anti-Acetyl-Histone H3 (Lys18) antibody (PTMBIO, #PTM-114RM, 1:1000), anti-Acetyllysine antibody (PTMBIO, #PTM-101, 1:1000), and anti-Acetyl-Histone H4 (Lys8) antibody (PTMBIO, #PTM-190, 1:1000).

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 primary antibodies used here included anti-LDHA antibody (Proteintech, #19987-1-AP, 1:3000), anti-LDHB antibody (Proteintech, #14824-1-AP, 1:5000), anti-H3 antibody (Proteintech, #17168-1-Ap, 1:3000), anti-PCAF antibody (Cell Signaling Technology, #3378S, 1:1000), anti-P300 antibody (Proteintech, #20695-1-AP, 1:500), anti-GCN5 antibody (Proteintech, #66575-1-Ig, 1:3000), anti-beta Actin antibody (PTMBIO, #PTM-5028, 1:1000), anti-human CXCL1 Polyclonal antibody (Proteintech, #12335-1-AP), anti-mouse CXCL1 Polyclonal antibody (Proteintech, #30322-1-AP), anti-L-Lactyl Lysine antibody (PTMBIO, #PTM-1401RM, 1:1000), anti-L-Lactyl-Histone H3K18 antibody (PTMBIO, #PTM-1406RM, 1:1000), anti-L-Lactyl-Histone H3K9 antibody (PTMBIO, #PTM-1419RM, 1:1000), anti-L-Lactyl-Histone H3K14 antibody (PTMBIO, #PTM-1414RM, 1:1000), anti-H4 antibody (PTMBIO, #PTM-1009, 1:1000), anti-L-Lactyl-Histone H4K12 antibody (PTMBIO, #PTM-1411RM, 1:1000), anti-L-Lactyl-Histone H4K8 antibody (PTMBIO, #PTM-1415RM, 1:1000), anti-Acetyl-Histone H3 (Lys18) antibody (PTMBIO, #PTM-114RM, 1:1000), anti-Acetyllysine antibody (PTMBIO, #PTM-101, 1:1000), and anti-Acetyl-Histone H4 (Lys8) antibody (PTMBIO, #PTM-190, 1:1000).

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