mouse ccl2 je mcp 1  (R&D Systems)

Journal: Redox Biology

Article Title: The electrophilic metabolite of kynurenine, kynurenine-CKA, requires C151 in Keap1 to derepress Nrf2

doi: 10.1016/j.redox.2026.104009

Figure Lengend Snippet: AhR inhibition has no effect on the anti-inflammatory actions of Kyn-CKA in BMDMs. (A) Mouse AhR reporter cells expressing luciferase under the control of the xenobiotic response element (XRE) were incubated with Kyn-CKA or kynurenine for 24 h prior to luminescence measurement (n = 3). Data were fitted to a sigmoidal four-parameter logistic curve (L-Kyn: r 2 = 0.824, IC 50 = 28 μM, span = 3218 RFU. Kyn-CKA: r 2 = 0.970, IC 50 = 13 μM, span = 47,901 RFU). (B) Inhibition of AhR-dependent luciferase expression by CH223191 (AhRinh). (C–F) Kyn-CKA inhibits the expression of the NF-κB-regulated genes IL6, MCP1, Nos2 and IL1β in BMDM following 5 h incubation with LPS (0.5 μg/mL) and the indicated concentrations of Kyn-CKA both in the presence or absence of CH223191 (10 μM). Data are n = 3–4, ∗∗∗∗p < 0.0001 by one-way ANOVA and Tukey's post-test. (G – H) Extracellular cytokine levels from BMDM treated as in C–F for 5 h. Data are n = 3–4 (all replicates shown), ∗∗∗∗p < 0.0001 by one-way ANOVA and Tukey's post-test.

Article Snippet: ELISA kits for MCP1 (DY479 and MJE00B) and IL6 (DY406) were obtained from R&D Systems (Minneapolis, MN).

Techniques: Inhibition, Expressing, Luciferase, Control, Incubation

Journal: Redox Biology

Article Title: The electrophilic metabolite of kynurenine, kynurenine-CKA, requires C151 in Keap1 to derepress Nrf2

doi: 10.1016/j.redox.2026.104009

Figure Lengend Snippet: The low-dose anti-inflammatory effects of Kyn-CKA in BMDMs are dependent on Nrf2. (A – B) Treatment with Kyn-CKA (5 h) fails to induce the expression of Nqo1 and Gclm in BMDMs obtained from Nrf2-knockout (Nrf2 −/− ) mice. Data are n = 3, ∗∗∗∗p < 0.0001 by two-way ANOVA and Tukey's post-test. (C – G) Effects of Kyn-CKA on the expression of NF-κB−regulated genes in WT and Nrf2-KO BMDMs following 5 h incubation with LPS (0.5 μg/mL) and the indicated concentrations of Kyn-CKA. Data are n = 6, ∗p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 by two-way ANOVA and Tukey's post-test. Nrf2 was identified as a significant source of variation for MCP1, IL6, TNFα, Nos2, (p < 0.0001) and IL1β (p < 0.005).

Article Snippet: ELISA kits for MCP1 (DY479 and MJE00B) and IL6 (DY406) were obtained from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Knock-Out, Incubation

Journal: Nature Communications

Article Title: Activation of IRF3 in cardiomyocytes impairs mitochondrial oxidative function through PGC-1α inhibition and drives heart failure

doi: 10.1038/s41467-026-69792-4

Figure Lengend Snippet: a Immunoblot showing expression of p-IRF3 and IRF3 levels in the left ventricle tissue of humans with ischemic cardiomyopathy (ICM) compared to non-failing (NF) hearts, NF ( n = 6) and ICM ( n = 6), biological replicates. Vinculin was used as a loading control. b Quantification of the immunoblot shown in Fig. 1a. Statistical significance was calculated by unpaired two-tailed Student’s t test, NF ( n = 6) and ICM ( n = 6), *** P = 5.1 × 10 -7 . c mRNA expression of IRF3 and target genes in the left ventricle tissue of human patients with ICM compared to NF hearts determined by qPCR. Statistical significance was calculated by unpaired two-tailed Student’s t test, samples from NF ( n = 7) and ICM patients ( n = 10) are biological replicates, IRF3 : ** P = 0 0.0230; IFIT2 : * P = 0.0534. d UMAP showing single-cell analysis of genes regulated in the left ventricle tissue of patients with heart failure compared to healthy control hearts. e Gene expression of IRF3 in cardiomyocytes isolated from patients with heart failure. f Immunoblot showing expression of p-IRF3 and IRF3 levels in the left ventricle tissue of mice with LAD induced myocardial infarction (MI) compared to Sham operated αMHC Cre control. Vinculin was used as a loading control. N = 3 per group, biological replicates. g Box plot showing gene expression of Irf3 and target genes regulating type I IFN signaling in the left ventricle tissue of mice with LAD induced MI determined by qPCR. Sham ( n = 7) and LAD ( n = 8), biological replicates. Box and whiskers plot showing all minimum to maximum points. Significance was calculated by unpaired two-tailed Student’s t test, Irf3 : ** P = 0.0047; Ifnβ : ** P = 0.0041; Ifit1 : ** P = 0.0022; Ifit2 : ** P = 0.0099; Ifit3 : ** P = 0.0046; Rsad2 : * P = 0.0438; Ccl2: * P = 0.0311. h Immunoblot showing expression of OXPHOS proteins [NADH: ubiquinone oxidoreductase (NDUFB8), Succinate dehydrogenase (SDHB), Cytochrome c oxidase I (CoI), Ubiquinol Cytochrome c oxidoreductase (UQCRC2), ATP synthase (ATP5A)] in the left ventricle tissue of humans with ICM compared to NF hearts, n = 6 per group, biological replicates. i Quantification of the immunoblot shown in Fig. 1h. Statistical significance was calculated by unpaired two-tailed Student’s t test, n = 6 per group, biological replicates, NDUFB8: *** P = 0.0005; SDHB: * P = 0.0269; UQCRC2: * P = 0.0162; COI: ** P = 0.0024; ATP5A: ** P = 0.0015. j Gene expression of mitochondrial OXPHOS marker genes by qPCR in the left ventricle tissue of mice with LAD induced MI. Box and whiskers plot showing all minimum to maximum points. Sham ( n = 7) and LAD ( n = 8), biological replicates. Significance was calculated by unpaired two-tailed Student’s t test and P values are shown in the box plot. k Nd1 , 16sRNA (mitochondrial) and Sdhb (nuclear) levels in mtDNA isolated from rat left ventricle was quantified by qPCR. Data points represent mtDNA samples ( n = 4), biological replicates. l Immunoblot showing pIRF3 levels in primary cardiomyocytes treated with 1 µg/ml of freshly isolated mtDNA for 6 h. N = 3 independent replicates per group. Data in all panels are represented as mean ± SEM. Source data are provided as a Source file.

Article Snippet: The concentration of TNF-α (#DY410), CCL2 (#DY479), IL-1β (#DY401), and IL-6 (#DY406) was measured in the mouse plasma samples using the equivalent DuoSet ELISA kits (R&D Systems, Minneapolis, MN, USA) following the manufacturer’s protocols.

Techniques: Western Blot, Expressing, Control, Two Tailed Test, Single-cell Analysis, Gene Expression, Isolation, Marker

Journal: Nature Communications

Article Title: Activation of IRF3 in cardiomyocytes impairs mitochondrial oxidative function through PGC-1α inhibition and drives heart failure

doi: 10.1038/s41467-026-69792-4

Figure Lengend Snippet: a Representative model showing strategy of CMI3OE transgenic mouse generation. Irf3-2D fl/fl mice were bred with αMHCMerCreMer (αMHCMCM) mice to generate tamoxifen inducible cardiomyocyte-specific CMI3OE ( Irf3-2D fl/αMHCMCM) mice. b Schematic representation of cardiomyocyte-specific expression of murine IRF3-2D by oral gavage of low dose tamoxifen (30 mg/kg) to induce MerCreMer-mediated excision of cardiomyocyte-specific floxed cassette in 12wk old male mice. c Gross morphology of αMHCMCM (Cre) and CMI3OE male mice at 12wks of age. d Kaplan-Meier curve showing survival of CMI3OE compared to αMHCMCM (Cre) mice. e Western blot showing protein levels of IRF3 in the cardiac ventricular tissue, n = 3 per group, biological replicates. f Quantification of the Western blot shown in Fig. 3e, n = 3 per group, biological replicates. Data analyzed by unpaired two-tailed Student’s t test ***P = 2 × 10 -5 . g mRNA expression of Irf3 and its target genes in the cardiac ventricular tissue determined by qPCR, n = 7 per group, biological replicates. Data analyzed by unpaired two-tailed Student’s t test, Irf3 : *** P = 1.8 × 10 -8 ; Ifnβ : * P < 0.0280; Ifit1 : *** P < 1.5 × 10 -9 ; Ifit2 : *** P < 0.0005; Ifit3 : *** P < 0.0002; Rsad2 : *** P < 9.9 × 10 -7 ; Ccl2 : ** P < 0.0030. h-j Plasma cytokine levels in CMI3OE mice determined by ELISA. Samples used are biological replicates. Data analyzed by unpaired two-tailed Student’s t test, CCL2: * P = 0.0430 [ n = 5 per group]; TNFα: * P = 0.0229 [ n = 5 per group]; IL-6: * P = 0.0532 [ n = 6 (Cre), n = 9 (CMI3OE)]. k Body weight measured in αMHCMCM (Cre) and CMI3OE mice, n = 5 per group. l Representative echocardiography images from αMHCMCM (Cre) and CMI3OE mice. m – q Cardiac function in αMHCMCM (Cre) and CMI3OE mice determined by echocardiography, n = 5 per group, biological replicates. Data analyzed by unpaired two-tailed Student’s t test, %EF: *** P = 2.3 × 10 -5 ; %FS: *** P = 5.8 × 10 -5 ; LVPWd: * P = 0.0448; LVmass/BW: * P = 0.0533. Cardiac function was measured in three independent cohorts with similar results. r Heart weight to tibia length determined in αMHCMCM (Cre) and CMI3OE mice, n = 5 per group, data analyzed by unpaired two-tailed Student’s t test, * P = 0.0399. s Lung weight to tibia length determined in αMHCMCM (Cre) and CMI3OE mice, n = 5 per group. Survival curve, gene expression analysis, and echocardiography experiments were performed using independent cohorts. Data in all panels are represented as mean ± SEM. Source data are provided as a Source data file.

Article Snippet: The concentration of TNF-α (#DY410), CCL2 (#DY479), IL-1β (#DY401), and IL-6 (#DY406) was measured in the mouse plasma samples using the equivalent DuoSet ELISA kits (R&D Systems, Minneapolis, MN, USA) following the manufacturer’s protocols.

Techniques: Transgenic Assay, Expressing, Western Blot, Two Tailed Test, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Gene Expression