Journal: CNS Neuroscience & Therapeutics

Article Title: Histone H3K18 Lactylation Contributes to Perioperative Neurocognitive Disorder Through Immune Checkpoint Lymphocyte Activation Gene 3 Mediated Microglial Pyroptosis

doi: 10.1002/cns.71058

Figure Lengend Snippet: The target genes regulated by H3K18la were identified by CUT&Tag assay combined with RNA‐seq. (A) Average plot and heat map displaying the binding density of H3K18la with different H3K18la binding peaks in hippocampus of the Sur group and Con group. (B) Genome‐wide distribution of the upregulated H3K18la‐binding peaks in mice after surgery. (C) Bar graph illustrating the number of target genes associated with upregulated and downregulated H3K18la peaks, and the top 2 enriched de novo motifs of target genes. (D) GO analysis of candidate target genes associated with upregulated H3K18la peaks. (E) Volcano plot of differentially expressed genes in RNA‐seq ( n = 3). (F) Heat map showing TOP 20 differential genes after 2‐DG treatment in RNA‐seq ( n = 3). (G) Venn diagram showing the intersection of H3K18la up‐regulated target genes and 2‐DG down‐regulated target genes. (H) Genome browser tracks show H3K18la binding at representative target gene loci. Peaks specifically located at the Lag3 promoters are identified by red rectangles. (I) H3K18la occupancy analysis by ChIP‐qPCR ( n = 3). Data are mean ± SD. Statistical significance was assessed by unpaired 2‐tailed Student t ‐test compared with Con group. **** p < 0.0001.

Article Snippet: Sur + 2‐DG mice received 2‐DG (250 mg/kg; MedChemExpress, HY‐13966) [ ] intraperitoneally once daily for 2 days preoperatively until 30 min before surgery.

Techniques: RNA Sequencing, Binding Assay, Genome Wide, ChIP-qPCR

Journal: CNS Neuroscience & Therapeutics

Article Title: Histone H3K18 Lactylation Contributes to Perioperative Neurocognitive Disorder Through Immune Checkpoint Lymphocyte Activation Gene 3 Mediated Microglial Pyroptosis

doi: 10.1002/cns.71058

Figure Lengend Snippet: H3K18la promotes Lag3 expression in microglial cells induced by either lactate or LPS and IFN‐γ. (A) BV2 cells were first treated with 0‐, 1‐, 5‐, and 25‐mM lactate for 24 h. Subsequently, cells pretreated with 25 mM lactate or 10 mM 2‐DG for 3 h were stimulated with LPS and IFN‐γ for 24 h. (B) Representative immunoblotting of Pan‐Kla, H3K18la and Lag3 in BV2 cells intervention with lactate, with histone H3 or GAPDH used for normalization. (C‐E) Quantitation of Pan‐Kla, H3K18la and Lag3 gray value ( n = 3). (F) The lactate levels of BV2 cells activated by LPS and IFN‐γ ( n = 5–6). (G) Representative immunoblotting of Pan‐Kla, H3K18la and Lag3 in BV2 cells activated by LPS and IFN‐γ, with histone H3 or GAPDH used for normalization. (H–J) Quantitation of Pan‐Kla, H3K18la and Lag3 gray value ( n = 3–4). (K) Representative fluorescence images of Pan‐Kla or Lag3 in BV2 cells. (L, M) The relative fluorescence intensities of Pan and Lag3 in BV2 cells ( n = 6). The images include scale indicators of 50 μm for size reference. The data are presented as the mean ± SD, A–D were assessed by unpaired 2‐tailed Student t ‐test compared with Con group. The other results were assessed by one‐way ANOVA followed by Tukey's post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Sur + 2‐DG mice received 2‐DG (250 mg/kg; MedChemExpress, HY‐13966) [ ] intraperitoneally once daily for 2 days preoperatively until 30 min before surgery.

Techniques: Expressing, Western Blot, Quantitation Assay, Fluorescence