nos inhibitor trim  (MedChemExpress)

Journal: Nature Communications

Article Title: Exercise-induced histone lactylation in monocyte-derived macrophages restores cardiac immune homeostasis and function in sepsis-induced cardiomyopathy

doi: 10.1038/s41467-025-67443-8

Figure Lengend Snippet: Schematic representation of the mice treatment ( A ) and average voluntary running distance (km/day) ( B ). The analysis includes a total of 14 mice. Figure created in BioRender, Shang, M. (2025) tc3zfga. Body temperature ( C ) and percent change in body weight ( D ) of 18 h post intraperitoneal injection of PBS or LPS in sedentary (Sed; n = 8 for PBS, n = 6 for LPS) and exercise (Exe; n = 8 for PBS, n = 6 for LPS) mice. Representative echocardiography images ( E ) and quantification of EF% ( F ) and FS% ( G ) on 18 h post PBS or LPS i.p injection on Sed (n = 8 for PBS, n = 6 for LPS) and Exe (n = 8 for PBS, n = 6 for LPS) mice. Representative images and quantification of TUNEL ( H, J ) and DHE ( I, K ) in heart sections 18 h post intraperitoneal injection of PBS or LPS in Sed (n = 5 for PBS, n = 5 for LPS) and Exe (n = 5 for PBS, n = 5 for LPS) mice. The experiment shows representative values from 3 independent experiments. L Principal component analysis (PCA) of the transcriptomes of PBS (Sed), PBS (Exe), LPS (Sed), and LPS (Exe) groups. M Upset plot displays the number and overlap of DEGs between LPS (Sed) and PBS (Sed), LPS (Exe) and PBS (Exe), LPS (Exe) and LPS (Sed), PBS (Exe) and PBS (Sed) groups. N Heatmap of differential expression patterns. Samples from different groups are illustrated above, and representative genes with associated GO terms are illustrated. ELISA analysis of IL-1β ( O ) and TNF-α ( P ) in Sed and Exe mice. The analysis was performed on 6 samples per group. Quantitative of infiltrated macrophages (CD68 + ) ( R ) and CD68 + Arg1 + cells ( S ), and representative images of immunofluorescent staining of CD68 (green) and Arg1 (red) in hearts of mice ( Q ) with indicated treatments. The analysis was performed on 5 samples per group from 3 independent experiments. Data are represented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t-tests ( C, D, F–I, O, P, R, S ). NS, not significant. Scale bar = 100 µm ( J, K, Q ). Source data for B–D, F–I, O , P , R , and S are provided in the file.

Article Snippet: PEG-DSPE (20 mg), DSPE-PEG-NHS (20 mg), and the NOS inhibitor TRIM (26 mg) or Arg1 inhibitor piceatannol 3’-O-glucoside (HY-N2237, MCE) were dissolved in 2 mL tetrahydrofuran and added dropwise to 5 mL PBS solution while stirring for 2 h. After dialyzing the solution in PBS for 24 h (MWCO = 3500), Ly6c antibody (1 mg) was added and stirred for an additional 12 h, followed by dialysis in PBS for another 24 h (MWCO = 30,000).

Techniques: Injection, TUNEL Assay, Quantitative Proteomics, Enzyme-linked Immunosorbent Assay, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Exercise-induced histone lactylation in monocyte-derived macrophages restores cardiac immune homeostasis and function in sepsis-induced cardiomyopathy

doi: 10.1038/s41467-025-67443-8

Figure Lengend Snippet: A The Monocle prediction of the monocyte-macrophage developmental trajectory for Ly6c2 high Mo, Ccr2 + Mø, MHC II high Mø and iNOS + Mo. B Heatmap displaying differentially expressed genes based on their shared kinetics through pseudotime using Monocle2. Distinct GO terms and representative genes associated with each model are shown on the left and right. C Schematic illustration of the nano-inhibitor designed to specifically inhibit iNOS in monocytes. Figure created in BioRender, Shang, M. (2025) lbdglen. D , E Efficiency of iNOS inhibition by the nano-inhibitor in monocytes, detected by WB. β-Tubulin was used as loading control. Numbers represent densitometric fold change relative to β-Tubulin. The analysis was performed on 6 samples per group. Body temperature ( F ) and percent change in body weight ( G ) measured 18 h after i.p. injection of LPS with Anti-Ly6C-iNOS or control in sedentary (Sed; CTRL, n = 7; Anti-Ly6C-iNOS, n = 7) and exercise (Exe; CTRL, n = 8; Anti-Ly6C-iNOS, n = 7) mice. Representative echocardiography images ( H ) and quantification of EF% ( I ), FS % ( J ) at 18 h post-LPS i.p injection with Anti-Ly6C-iNOS or control in Sed (CTRL, n = 7; Anti-Ly6C-iNOS, n = 7) and Exe (CTRL, n = 8; Anti-Ly6C-iNOS, n = 7) mice. Representative images and quantification of TUNEL + cells ( N, K ), DHE + areas ( O, L ) and iNOS + CD68 + cells ( P, M ) in hearts of Sed and Exe mice 18 h post LPS injection, treated with either Anti-Ly6C-iNOS or control (CTRL). The analysis was performed on 4 samples per group from 2 independent experiments. Q–S Representative images and quantification of Arg1, CD68 and iNOS in hearts of mice over a time course following LPS injection. The analysis was performed on 5 samples per group from 3 independent experiments. Body temperature ( T ) and percent change in body weight ( U ) measured 18 h after i.p. injection of LPS with Anti-Ly6C-Arg1 or control in Sed and Exe mice. The analysis was performed on 6 samples per group. Representative echocardiography images ( V ) and quantification of EF% ( W ) and FS% ( X ) measured 18 h after i.p. injection of LPS with Anti-Ly6C-Arg1 or control in Sed and Exe mice. The analysis was performed on 6 samples per group. Data are represented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t-tests. NS not significant. Scale bar: 100 µm ( N – P ), 50 µm ( Q ). Source data for D – G, I – M, R – U and W – X are provided in the file.

Article Snippet: PEG-DSPE (20 mg), DSPE-PEG-NHS (20 mg), and the NOS inhibitor TRIM (26 mg) or Arg1 inhibitor piceatannol 3’-O-glucoside (HY-N2237, MCE) were dissolved in 2 mL tetrahydrofuran and added dropwise to 5 mL PBS solution while stirring for 2 h. After dialyzing the solution in PBS for 24 h (MWCO = 3500), Ly6c antibody (1 mg) was added and stirred for an additional 12 h, followed by dialysis in PBS for another 24 h (MWCO = 30,000).

Techniques: Inhibition, Control, Injection, TUNEL Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Exercise-induced histone lactylation in monocyte-derived macrophages restores cardiac immune homeostasis and function in sepsis-induced cardiomyopathy

doi: 10.1038/s41467-025-67443-8

Figure Lengend Snippet: A Glycolysis and TCA cycle activity scores in peripheral blood monocytes from healthy donors and SICM patients based on GSVA analysis. Number of samples included in the analysis: Control, n = 9; SICM, n = 10. The lower whisker, lower hinge, box center, upper hinge, and upper whisker represent the minimum, lower quartile, median, upper quartile. B Volcano plot showing differential gene expression between healthy donors with SICM patients, and LdhA is indicated in the graph. Body temperature ( C ) and percentage change in body weight ( D ) of myeloid specific Ldha deletion ( Ldha ΔMo ) and control mice 18 h post LPS injection, with or without exercise. Groups include sedentary (Sed; CTRL, n = 6; Ldha ΔMo , n = 4) and exercise (Exe; CTRL, n = 5; Ldha ΔMo , n = 7). Representative echocardiography images ( E ) and quantification of EF% ( F ) and FS % ( G ) in Ldha ΔMo or control mice 18 h post-LPS injection, with or without exercise. Groups include sedentary (CTRL, n = 6; Ldha ΔMo , n = 4) and exercise (CTRL, n = 5; Ldha ΔMo , n = 7). Representative images and quantification of TUNEL ( J , H ) and DHE ( K , I ) positive area in hearts of Ldha ΔMo or control mice 18 h post LPS injection, with or without exercise. The analysis was performed on 4 samples per group. L–P , Representative images and quantification of Pan Kla, iNOS, CD68 and Arg1 in the hearts of Ldha ΔMo or control mice 18 h post LPS injection, with or without exercise. The analysis was performed on 5 samples per group. All experiments except A and B show representative values from three independent experiments. Data are represented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t-tests, except for A, where unpaired Mann–Whitney was used. NS not significant. Scale bar: 100 µm ( J – L ). Source data for A – D , F – I and M – P are provided in the file.

Article Snippet: PEG-DSPE (20 mg), DSPE-PEG-NHS (20 mg), and the NOS inhibitor TRIM (26 mg) or Arg1 inhibitor piceatannol 3’-O-glucoside (HY-N2237, MCE) were dissolved in 2 mL tetrahydrofuran and added dropwise to 5 mL PBS solution while stirring for 2 h. After dialyzing the solution in PBS for 24 h (MWCO = 3500), Ly6c antibody (1 mg) was added and stirred for an additional 12 h, followed by dialysis in PBS for another 24 h (MWCO = 30,000).

Techniques: Activity Assay, Control, Whisker Assay, Gene Expression, Injection, TUNEL Assay, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Exercise-induced histone lactylation in monocyte-derived macrophages restores cardiac immune homeostasis and function in sepsis-induced cardiomyopathy

doi: 10.1038/s41467-025-67443-8

Figure Lengend Snippet: A – C Western blot for Pan Kla and H3K18la in mice monocytes incubated with 20 mM sodium lactate for 24 h. Histone H3 was used as an internal control, with densitometric fold change relative to Histone H3 indicated. The analysis was performed on 9 samples per group. Western blot for H3K18la in mouse monocytes subjected to p300 or HDAC2 knockdown using si-P300 ( D – F ) or si-HDAC2 ( G – I ), treatment with 20 mM sodium lactate for 24 h, and their respective control groups. Tubulin and Histone H3 were used as internal controls, with densitometric fold change relative to Tubulin and Histone H3 indicated. The analysis was performed on 3 samples per group. J Schematic presentation of BMDMs stimulated with LPS over a time course following incubation with 20 mM sodium lactate for 24 h. K , L , N Representative images and quantification of Arg1 and iNOS in lactate pre-incubated BMDMs upon LPS challenge. The analysis was performed on 4 samples per group. RT-qPCR analysis of Arg1 ( M ) and iNOS ( O ) in lactate pre-incubated BMDMs upon LPS challenge. The analysis was performed on 3 samples per group. Representative traces of CUT&Tag analysis showed that H3K18la was enriched in the promoter regions of Arg1 ( P ). CUT&Tag results for Arg1 were verified by qPCR ( Q ). The analysis was performed on 3 samples per group. Exercise is abbreviated as Exe and sedentary as Sed. All experiments show representative values from at least 2 independent experiments. Data are represented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t-tests. NS, not significant. Scale bar: 250 µm ( K ). Source data for B , C , E , F , H , I , L – O and Q are provided in the file.

Article Snippet: PEG-DSPE (20 mg), DSPE-PEG-NHS (20 mg), and the NOS inhibitor TRIM (26 mg) or Arg1 inhibitor piceatannol 3’-O-glucoside (HY-N2237, MCE) were dissolved in 2 mL tetrahydrofuran and added dropwise to 5 mL PBS solution while stirring for 2 h. After dialyzing the solution in PBS for 24 h (MWCO = 3500), Ly6c antibody (1 mg) was added and stirred for an additional 12 h, followed by dialysis in PBS for another 24 h (MWCO = 30,000).

Techniques: Western Blot, Incubation, Control, Knockdown, Quantitative RT-PCR, Two Tailed Test