foxo3  (Addgene inc)

Journal: The Korean Journal of Pain

Article Title: IL-27-Ucp2-FoxO3 axis mediating the polarization of alternatively activated macrophages and ameliorating inflammatory pain

doi: 10.3344/kjp.25307

Figure Lengend Snippet: A schematic overview presents the framework of this study. AAMs: alternatively activated macrophage, FoxO3: forkhead box class O3, IL: interleukin.

Article Snippet: FoxO3 (D19A7) Rabbit mAb , CST, US , #12829.

Techniques:

Journal: The Korean Journal of Pain

Article Title: IL-27-Ucp2-FoxO3 axis mediating the polarization of alternatively activated macrophages and ameliorating inflammatory pain

doi: 10.3344/kjp.25307

Figure Lengend Snippet: IL-27-Ucp2-mediated AAM and a subsequent activation of the transcription factor FoxO3. (A) A diagram of the experimental process of this section. (B) Volcano plot analysis of differentially expressed proteins (DEPs) comparing IL-27 vs. NC, IL-27 vs. si-Ucp2, and IL-27 vs. GNP-treated groups. The marked Acod1 was indicated. (C) IL-27 induced a higher expression of Irg1 in BMDM, n = 3, GAPDH was used as a reference, and an unpaired two-tailed t -test was applied. (D) Compared to the WT group, IL-27-induced Arg-1 expression was inhibited in the si-Irg1 treated group. GAPDH was used as a reference, n = 3; the P values (si-Irg1 vs. WT in NC/IL-27/IL-4 treatment) were tested by two-way ANOVA with Tukey’s multiple comparisons test. (E) Transcription factor (TF) prediction for both DEGs and DEPs induced by IL-27-stimulated BMDM. (F) The binding motif of FoxO3 predicted by the JASPAR platform. (G) The mRNA level of FoxO3 in BMDM with either NC, IL-27 (100 ng/mL), and IL-4 (20 ng/mL) treatment. N = 4, GAPDH was used as a reference, and an unpaired two-tailed t -test was applied. (H) Compared to the WT group, IL-27-induced FoxO3 expression was inhibited in si-Irg1/si-Ucp2/GNP-treated group. GAPDH was used as a reference, n = 4; two-way ANOVA tested the P values with Tukey’s multiple comparisons test. ** P < 0.01, *** P < 0.001. Data are shown as mean ± standard error of the mean. IL: interleukin, Ucp2: uncoupling protein 2, AAM: alternatively activated macrophage, GNP: Genipin explicitly inhibits the protein function of Ucp2, BMDM: bone marrow-derived macrophage, WT: wild type, DEGs: differential expression analysis of genes.

Article Snippet: FoxO3 (D19A7) Rabbit mAb , CST, US , #12829.

Techniques: Activation Assay, Expressing, Two Tailed Test, Binding Assay, Derivative Assay, Quantitative Proteomics

Journal: The Korean Journal of Pain

Article Title: IL-27-Ucp2-FoxO3 axis mediating the polarization of alternatively activated macrophages and ameliorating inflammatory pain

doi: 10.3344/kjp.25307

Figure Lengend Snippet: FoxO3 controls IL-27-induced AAM differentiation and mitigates inflammatory pain in mice. (A) Immunofluorescence (IF) microscopy of FoxO3 in the nucleus of BMDMs upon stimulation with either NC, IL-27 (100 ng/mL for 12 hours), and IL-4 (20 ng/mL for 24 hours). n = 3, The scale bar indicates 1cm, and (B) the P values were tested by a one-way ANOVA with Tukey’s multiple comparisons test. (C) By comparing the knockdown efficiencies of three candidate siRNAs targeting FoxO3, Si#1 was identified as the most effective and will use it in subsequent experiments. GAPDH was used as a reference, n = 3, and the P values ( vs. NC) were tested by a one-way ANOVA with Dunnett’s post hoc test. (D) Compared to the FoxO3 mRNA in WT and/or si-Fxox3-treated BMDM in response to IL-27 (100 ng/mL for 12 hours) and IL-4 (20 ng/mL for 24 hours) treatment. GAPDH was used as a reference, n = 4, and the P values (si-FoxO3 vs. WT in NC/IL-27/IL-4 treatment) were tested by two-way ANOVA with Tukey’s multiple comparisons test. (E–G) Three M2 markers (Arg-1, Chi3l3, Retnla) were assessed by qPCR (E) and ELISA methods (F, G) in WT and/or si-Fxox3-treated BMDM in response to IL-27 (100 ng/mL for 12 hours) and IL-4 (20 ng/mL for 24 hours) treatment. GAPDH was used as a reference, n = 3, and the P values (si-FoxO3 vs. WT in NC/IL-27/IL-4 treatment) were tested by two-way ANOVA with Tukey’s multiple comparisons test. (H) Flowchart of the experimental process of the adoptive transfer strategy. (I, J) Course of mechanical hyperalgesia in mice with five different intervention groups. (I) The BMDM transfer time was indicated, n = 5, and P values (M vs. MSF3) were tested by two-way ANOVA with Tukey’s multiple comparisons test. (J) The results of IL-27M-IL-27MSF3 groups. N = 5, and P values (IL-27M vs. IL-27MSF3) were tested by two-way ANOVA with Tukey’s multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001, ns: not significant. Data are shown as mean ± standard error of the mean. FoxO3: forkhead box class O3, IL: interleukin, Ucp2: uncoupling protein 2, AAM: alternatively activated macrophage, BMDM: bone marrow-derived macrophage, WT: wild type, M: macrophages, MSF3: macrophage + si-FoxO3, IL-27M: IL-27-primed macrophage, IL-27MSF3: IL-27-primed macrophage+si-FoxO3.

Article Snippet: FoxO3 (D19A7) Rabbit mAb , CST, US , #12829.

Techniques: Immunofluorescence, Microscopy, Knockdown, Enzyme-linked Immunosorbent Assay, Adoptive Transfer Assay, Derivative Assay

Journal: Biochemistry and Biophysics Reports

Article Title: Mitigating psychological stress-induced cardiac injury: The impact of CB2R agonists on endoplasmic reticulum stress and mitophagy

doi: 10.1016/j.bbrep.2025.102425

Figure Lengend Snippet: Correlation analysis between key genes and immune-cell signatures. (A) Heatmap showing Pearson correlations between the expression of Foxo3, Pparg, Sirt1, and Stat3 and ssGSEA-derived immune-cell scores (red, positive; blue, negative). (B–D) Scatter plots showing positive correlations between Sirt1 expression and CD56 ∧ bright natural killer cells (B), myeloid-derived suppressor cells (MDSCs) (C), and type 1 T helper (Th1) cells (D). (E) Scatter plot showing a positive correlation between Pparg expression and Th1 cells. (F) Scatter plots showing negative correlations between γδ T cells and Stat3, Sirt1, Pparg, and Foxo3. Pearson's r and P values are indicated in each panel.

Article Snippet: Protein expression levels of FOXO3, STAT3, PPARγ, and SIRT1 were analyzed using the following primary antibodies: PPARγ (Abcam, UK, ab178860), FOXO3 (Cell Signaling Technology, USA, #12829), STAT3 (Abclonal, China, A22434), and SIRT1 (Abclonal, China, A0230).

Techniques: Expressing, Derivative Assay

Journal: Biochemistry and Biophysics Reports

Article Title: Mitigating psychological stress-induced cardiac injury: The impact of CB2R agonists on endoplasmic reticulum stress and mitophagy

doi: 10.1016/j.bbrep.2025.102425

Figure Lengend Snippet: Analysis of Key Genes . (A) Comparison of key gene expression between the stress and treatment groups. The x-axis represents the four key genes, while the y-axis indicates gene expression levels. Red denotes the stress group, and yellow denotes the treatment group. (B) Comparison of key gene expression in the validation set ( GSE68077 ) between controls and 5dayStress-1dayRest samples. The x-axis represents the key genes, while the y-axis indicates gene expression levels. Blue denotes the control group, and red denotes the 5dayStress-1dayRest group. (C–F) ROC curves for the validation set, showing the diagnostic performance of Stat3 (C), Foxo3 (D), Pparg (E), and Sirt1 (F). The x-axis represents 1-Specificity, and the y-axis represents Sensitivity. The area under the curve (AUC) is displayed in the bottom right corner of each plot. (G–I) ROC curves for phenotype scores in GSE68077 (controls vs 5dayStress-1dayRest; G), test set control vs stress (H), and test set stress vs CB2R-agonist treatment (I). The x-axis represents 1-Specificity, and the y-axis represents Sensitivity. The area under the curve (AUC) is annotated in the bottom right corner of each plot. An AUC between 0.5 and 0.7 suggests low accuracy, between 0.7 and 0.9 suggests moderate accuracy, and >0.9 suggests high accuracy. ROC: Receiver Operating Characteristic. AUC: Area Under Curve. PTSD: Post-Traumatic Stress Disorder.

Article Snippet: Protein expression levels of FOXO3, STAT3, PPARγ, and SIRT1 were analyzed using the following primary antibodies: PPARγ (Abcam, UK, ab178860), FOXO3 (Cell Signaling Technology, USA, #12829), STAT3 (Abclonal, China, A22434), and SIRT1 (Abclonal, China, A0230).

Techniques: Comparison, Gene Expression, Biomarker Discovery, Control, Diagnostic Assay

Journal: Biochemistry and Biophysics Reports

Article Title: Mitigating psychological stress-induced cardiac injury: The impact of CB2R agonists on endoplasmic reticulum stress and mitophagy

doi: 10.1016/j.bbrep.2025.102425

Figure Lengend Snippet: Validation of Four Key Genes . (A–D) Relative mRNA expression levels of Sirt1 (A), Foxo3 (B), Pparg (C), and Stat3 (D) measured by quantitative real-time PCR (qPCR). Each qPCR analysis was conducted using cardiac tissue from six mice per group. (E) Protein expression levels of FOXO3, PPARG, SIRT1, and STAT3 as determined by Western blot analysis, with three biological replicates per group. Data are presented as mean ± SEM. Statistical significance is indicated as follows: ∗∗ (p < 0.01), ∗∗∗ (p < 0.001), ∗∗∗∗ (p < 0.0001).

Article Snippet: Protein expression levels of FOXO3, STAT3, PPARγ, and SIRT1 were analyzed using the following primary antibodies: PPARγ (Abcam, UK, ab178860), FOXO3 (Cell Signaling Technology, USA, #12829), STAT3 (Abclonal, China, A22434), and SIRT1 (Abclonal, China, A0230).

Techniques: Biomarker Discovery, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Mitigating psychological stress-induced cardiac injury: The impact of CB2R agonists on endoplasmic reticulum stress and mitophagy

doi: 10.1016/j.bbrep.2025.102425

Figure Lengend Snippet: Schematic model of chronic psychological stress–induced cardiac injury and the putative protective effect of CB2 receptor agonism. Chronic psychological stress triggers neurohumoral activation (sympathetic nervous system, HPA axis, and Ang II), promoting cardiomyocyte endoplasmic reticulum (ER) stress characterized by Ca 2+ overload and increased ROS, together with mitochondrial dysfunction. Within cardiomyocytes, four key nodes identified in this study (FOXO3, PPARγ, SIRT1, and STAT3) are depicted as an interacting regulatory network linking stress/inflammatory cues to mitochondrial quality control. SIRT1 activates FOXO3 via deacetylation, and FOXO3 induces BNIP3- and PINK1–PRKN-associated mitophagy gene programs, facilitating autophagosomal engulfment of damaged mitochondria; STAT3 shows bidirectional crosstalk with FOXO3 that may influence FOXO3 localization and transcriptional output in a cytokine-responsive context. PPARγ is illustrated as a regulator of immunometabolic remodeling and mitochondrial turnover pathways, including PGC-1α-linked metabolic reprogramming and FUNDC1-associated mitophagy. Mitophagy is presented as a set-point process with potentially adaptive versus excessive activation. Surrounding immune-cell alterations inferred from immune-signature analyses are shown, including increased CD56 ∧ bright NK cells, MDSCs, and Th1 cells, and decreased γδ T cells, accompanied by cytokine-driven signaling (e.g., IL-6, TNF) feeding into STAT3 and NF-κB/inflammatory pathways. Activation of cardiac CB2R by the agonist AM1241 is proposed to dampen ER stress/ROS and inflammatory signaling (NF-κB/JAK–STAT), restrain excessive mitophagy, normalize FOXO3/PPARγ/SIRT1/STAT3 expression, and reduce pro-inflammatory immune-cell infiltration, thereby promoting mitochondrial homeostasis. Green arrows indicate activation, and red dashed bar-headed lines indicate inhibition; upward/downward markers denote relative increases/decreases observed in this study. Abbreviations: Ang II, angiotensin II; CB2R, cannabinoid receptor type 2; ER, endoplasmic reticulum; HPA, hypothalamic–pituitary–adrenal; JAK, Janus kinase; MDSC, myeloid-derived suppressor cell; NF-κB, nuclear factor kappa B; PRKN, Parkin; ROS, reactive oxygen species; SNS, sympathetic nervous system.

Article Snippet: Protein expression levels of FOXO3, STAT3, PPARγ, and SIRT1 were analyzed using the following primary antibodies: PPARγ (Abcam, UK, ab178860), FOXO3 (Cell Signaling Technology, USA, #12829), STAT3 (Abclonal, China, A22434), and SIRT1 (Abclonal, China, A0230).

Techniques: Activation Assay, Control, Expressing, Inhibition, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Paederoside Promotes Longevity and Fitness in C. elegans Through Ubiquitination and Degradation of DAF-2/IGF1R, Activating DAF-16/FOXO and SKN-1/NRF2 Transcription Factors

doi: 10.3390/ijms27052248

Figure Lengend Snippet: ( a ) Molecular docking outcomes. Western blot analysis of ( b ) ubiquitination, ( c ) IGF1R, ( d ) CoIP and ( f ) key proteins of the IIS pathway. ( e ) FOXO3 nuclear translocation assay. The levels of statistical significance were set at * p < 0.05, ** p < 0.01, and **** p < 0.0001, ns and nd indicates no statistical significance.

Article Snippet: The antibodies used in this study were as follows: β-actin (HUABIO, Hangzhou, China, 1:20,000, HA722023); β-Tubulin (HUABIO, Hangzhou, China, 1:20,000, EM0103); IGF1R (abcam, Cambridge, MA, USA, 1:1000, ab182408, d,f); IGF1R (Proteintech Group, Wuhan, China, 1:1000, 21707-1-AP, c); ubiquitin (PTM bio, Hangzhou, China, 1:1000, PTM-7228); AKT3 (Absea Biotechnology, Suzhou, China, 1:1000, OC475); pAKT (Absea Biotechnology, Suzhou, China, 1:1000, RC4352); FOXO3 (abmart, Cambridge, MA, USA, 1:1000, PA5461S); and anti-Rabbit IgG-HRP (HUABIO, Hangzhou, China, 1:20,000, HA1001).

Techniques: Western Blot, Ubiquitin Proteomics, Nuclear Translocation Assay