acetyl foxo1  (Thermo Fisher)

Journal: Molecular Neurobiology

Article Title: SirT2 Inhibition is Associated with Improvements in Depression-like Behavior and Memory Impairment in Olfactory Bulbectomized Mice

doi: 10.1007/s12035-026-05868-y

Figure Lengend Snippet: Effects of AK-7 on the SirT2/Ac-FoxO1/PPARγ signaling pathway in the PFC of OBX mice. A - F Expression levels of SirT2 ( A ), Ac-FoxO1 ( B ), p-FoxO1 ( C ), PPARγ ( D ), p-AKT (Thr308) ( E ), and p-AKT (Ser473) ( F ) in the PFC of OBX mice. G: Representative immunofluorescence images showing spatial overlap of SirT2, Ac-FoxO1, and PPARγ with the microglial marker Iba1 in the PFC of OBX mice treated with AK-7. H: Proposed mechanistic model of AK-7 action. A – F One-way ANOVA; A F (2, 24) = 8.965, p = 0.0012; B F (2, 24) = 7.719, p = 0.0026; C F (2, 24) = 5.336, p = 0.0121; D F (2, 24) = 8.487, p = 0.0016; E F (2, 18) = 6.321, p = 0.0083; F F (2, 18) = 13.48, p = 0.0003. All data are presented as mean ± SEM. *: p < 0.05, **: p < 0.01 vs. sham + vehicle group. #: p < 0.05, ##: p < 0.01 vs. OBX + vehicle group

Article Snippet: Membranes were blocked for 30 min in Tris-buffered saline containing 0.01% Tween-20 (TBST) supplemented with 5% skim milk and subsequently incubated overnight at 4 °C with primary antibodies against SirT1 (1:2000; Millipore, Billerica, MA, USA; 07–131), SirT2 (1:2000; Cell Signaling Technology [CST], Danvers, MA, USA; #12650), SirT3 (1:1000; CST; #5490), SirT4 (1:2000; Proteintech, Rosemont, IL, USA; 66543–1-Ig), SirT5 (1:1000; CST; #8782), SirT6 (1:1000; CST; #12486), SirT7 (1:1000; CST; #5360), ΔFosB (1:500; CST; #14695), HRD1 (1:500; Proteintech; 13473–1-AP), acetyl-FoxO1 (Lys294; 1:200; Invitrogen, Waltham, MA, USA; PA5-104560), phospho-FoxO1 (Ser256; 1:500; CST; #9461), total FoxO1 (1:500; CST; #2880), PPARγ (1:500; CST; #2435), phospho-AKT (Thr308; 1:200; CST; #13038), phospho-AKT (Ser473; 1:1000; CST; #4060), total AKT (1:1000; CST; #4691), CD86 (1:500; Proteintech; 13395–1-AP), inducible nitric oxide synthase (iNOS; 1:50000; Proteintech; 18985–1-AP), interleukin-1β (IL-1β; 1:500; Proteintech; 26048–1-AP), tumor necrosis factor-α (TNF-α; 1:100; Proteintech; 17590–1-AP), arginase-1 (1:500; Proteintech; 66129–1-Ig), transforming growth factor-β (TGF-β; 1:500; Proteintech; 21898–1-AP), myelin basic protein (MBP; 1:1000; CST; #78896), myelin-associated glycoprotein (MAG; 1:1000; CST; #9043), cyclicnucleotide phosphodiesterase (CNPase; 1:1000; CST; #5664), contactin-associated protein (Caspr; 1:1000; Abcam, Cambridge, UK; ab246501), and β-actin (1:1000; Santa Cruz Biotechnology, Dallas, TX, USA; SC-47778).

Techniques: Expressing, Immunofluorescence, Marker

Journal: bioRxiv

Article Title: MATERNAL OBESITY MODULATES FOXO1 ACTIVATION AND ADIPOGENESIS IN NEONATAL MESENCHYMAL STEM CELLS

doi: 10.1101/2025.08.01.668117

Figure Lengend Snippet: A . Representative blot of expression of FOXO1, acetyl-FOXO1 and SIRT2 for both groups. B. FOXO1, acetyl-FOXO1, ratio of acetyl-FOXO1/total FOXO1 and SIRT2 on day 0. *p<0.05, Mann-Whitney U test (median ± range, n=8).

Article Snippet: After blocking, the membranes were incubated overnight at 4°C with antibodies for FOXO1 (Invitrogen, 1:500), acetyl-FOXO1 Lys 294 (Invitrogen, 1:1000), SIRT2 (Cell Signaling, 1:500), PPARγ (Cell Signaling, 1:250) and β-actin (Cell Signaling, 1:10000) and α-tubulin (Proteintech, 1:1000).

Techniques: Expressing, MANN-WHITNEY

Journal: bioRxiv

Article Title: MATERNAL OBESITY MODULATES FOXO1 ACTIVATION AND ADIPOGENESIS IN NEONATAL MESENCHYMAL STEM CELLS

doi: 10.1101/2025.08.01.668117

Figure Lengend Snippet: Cells were cultured with adipogenic induction media for 0, 1, 2, 3, 4 or 5 days and the expression of (A) FOXO1, (B) acetyl-FOXO1 and (C) SIRT2 was measured by western blot. Left: Representative western blots of NW-MSCs and OB-MSCs. Center: Levels of proteins over time in NW-MSCs and OB-MSCs. Right: Area under the curve (AUC) of levels of proteins in NW-MSCs vs OB-MSCs (median ± range, n=4). # p<0.05 Paired Wilcoxon signed-rank test compared to day 0 from the same group; *p<0.05 Mann-Whitney U test AUC.

Article Snippet: After blocking, the membranes were incubated overnight at 4°C with antibodies for FOXO1 (Invitrogen, 1:500), acetyl-FOXO1 Lys 294 (Invitrogen, 1:1000), SIRT2 (Cell Signaling, 1:500), PPARγ (Cell Signaling, 1:250) and β-actin (Cell Signaling, 1:10000) and α-tubulin (Proteintech, 1:1000).

Techniques: Cell Culture, Expressing, Western Blot, MANN-WHITNEY

Journal: bioRxiv

Article Title: MATERNAL OBESITY MODULATES FOXO1 ACTIVATION AND ADIPOGENESIS IN NEONATAL MESENCHYMAL STEM CELLS

doi: 10.1101/2025.08.01.668117

Figure Lengend Snippet: Cells were cultured with adipogenic induction media for 3 and 5 days, to assess the effect of H₂O₂ (250 μM) on FOXO1, acetyl-FOXO1 and SIRT2 levels. NW-MSCs (top), OB-MSCs (bottom) (median ± range, n=6). *p<0.05, Paired Wilcoxon signed-rank test. Representative western blots are shown in the left side.

Article Snippet: After blocking, the membranes were incubated overnight at 4°C with antibodies for FOXO1 (Invitrogen, 1:500), acetyl-FOXO1 Lys 294 (Invitrogen, 1:1000), SIRT2 (Cell Signaling, 1:500), PPARγ (Cell Signaling, 1:250) and β-actin (Cell Signaling, 1:10000) and α-tubulin (Proteintech, 1:1000).

Techniques: Cell Culture, Western Blot

Journal: bioRxiv

Article Title: MATERNAL OBESITY MODULATES FOXO1 ACTIVATION AND ADIPOGENESIS IN NEONATAL MESENCHYMAL STEM CELLS

doi: 10.1101/2025.08.01.668117

Figure Lengend Snippet: Cells were cultured with adipogenic induction media for 3 and 5 days, to assess the effect of AGK2 (10 μM), a SIRT2 inhibitor, on FOXO1, acetyl-FOXO1 and SIRT2 levels. NW-MSCs (top), OB-MSCs (bottom) (Median ± range, n=6). *p<0.05, Paired Wilcoxon signed-rank test. Representative western blots are shown in (left side).

Article Snippet: After blocking, the membranes were incubated overnight at 4°C with antibodies for FOXO1 (Invitrogen, 1:500), acetyl-FOXO1 Lys 294 (Invitrogen, 1:1000), SIRT2 (Cell Signaling, 1:500), PPARγ (Cell Signaling, 1:250) and β-actin (Cell Signaling, 1:10000) and α-tubulin (Proteintech, 1:1000).

Techniques: Cell Culture, Western Blot

Journal: bioRxiv

Article Title: MATERNAL OBESITY MODULATES FOXO1 ACTIVATION AND ADIPOGENESIS IN NEONATAL MESENCHYMAL STEM CELLS

doi: 10.1101/2025.08.01.668117

Figure Lengend Snippet: A. NW-MSCs and OB-MSCs were cultured with adipogenic induction medium and stained for acetyl-FOXO1 on days 0, 2, 4 and 6 of adipogenesis. Mean fluorescence intensity (MFI) of nuclear-cytoplasmatic ratio was calculated for localization. B . Representative images of day 2 for NW-MSCs (top) and OB-MSCs (bottom) (median ± range, n=6). *p<0.05, Mann-Whitney U test.

Article Snippet: After blocking, the membranes were incubated overnight at 4°C with antibodies for FOXO1 (Invitrogen, 1:500), acetyl-FOXO1 Lys 294 (Invitrogen, 1:1000), SIRT2 (Cell Signaling, 1:500), PPARγ (Cell Signaling, 1:250) and β-actin (Cell Signaling, 1:10000) and α-tubulin (Proteintech, 1:1000).

Techniques: Cell Culture, Staining, Fluorescence, MANN-WHITNEY

Journal: bioRxiv

Article Title: MATERNAL OBESITY MODULATES FOXO1 ACTIVATION AND ADIPOGENESIS IN NEONATAL MESENCHYMAL STEM CELLS

doi: 10.1101/2025.08.01.668117

Figure Lengend Snippet:

Article Snippet: After blocking, the membranes were incubated overnight at 4°C with antibodies for FOXO1 (Invitrogen, 1:500), acetyl-FOXO1 Lys 294 (Invitrogen, 1:1000), SIRT2 (Cell Signaling, 1:500), PPARγ (Cell Signaling, 1:250) and β-actin (Cell Signaling, 1:10000) and α-tubulin (Proteintech, 1:1000).

Techniques:

Journal: bioRxiv

Article Title: MATERNAL OBESITY MODULATES FOXO1 ACTIVATION AND ADIPOGENESIS IN NEONATAL MESENCHYMAL STEM CELLS

doi: 10.1101/2025.08.01.668117

Figure Lengend Snippet: MSCs of neonates from mothers with obesity have higher levels of total FOXO1 and SIRT2, together with lower acetyl-FOXO1, which induces an early commitment in response to adipogenic induction. On early day 2 of adipogenesis, acetyl-FOXO1 localizes mainly in the cytoplasma for OB-MSCs, therefore releases PPARγ for adipogenic gene transcription. Finally, on day 21, in vitro adipocytes of OB-MSCs show higher levels of PPARG gene expression, compared to NW-MSCs. Created in BioRender.

Article Snippet: After blocking, the membranes were incubated overnight at 4°C with antibodies for FOXO1 (Invitrogen, 1:500), acetyl-FOXO1 Lys 294 (Invitrogen, 1:1000), SIRT2 (Cell Signaling, 1:500), PPARγ (Cell Signaling, 1:250) and β-actin (Cell Signaling, 1:10000) and α-tubulin (Proteintech, 1:1000).

Techniques: In Vitro, Gene Expression

Journal: The FASEB Journal

Article Title: Ghrelin alleviates intestinal ischemia–reperfusion injury by activating the GHSR‐1α/Sirt1/FOXO1 pathway

doi: 10.1096/fj.202302155rrr

Figure Lengend Snippet: FIGURE 4 Ghrelin attenuates apoptosis in vitro by activating the GHSR-1α/Sirt1/FOXO1 signaling pathway. (A) Western blotting analysis of the expression of GHSR-1α, Sirt1, ac-FOXO1, and FOXO1. (B) Quantification of the protein levels of GHSR-1α, Sirt1, and ac- FOXO1/FOXO1 in Caco-2 cells. (C) Western blotting analysis of Bcl-2, Bax, and cleaved caspase3 expression. (D) Quantification of the relative protein levels of Bcl-2/Bax and cleaved caspase3. (E) Flow cytometry was used to detect the apoptosis rate. (F) Statistical analysis of the apoptosis rate. Data are expressed as mean ± SEM (n = 6). *p < .05 versus N; #p < .05 versus HR, +p < .05 versus HR + G.

Article Snippet: Antibodies Source Dilution Company Catalog number Ghrelin Rabbit 1:1000 Abmart, China TD6389S GHSR- 1α Rabbit 1:1000 Abmart, China TD2794S Sirt1 Rabbit 1:1000 CST, USA 9475S p- AMPK Rabbit 1:1000 CST, USA 50081S AMPK Rabbit 1:1000 Beyotime, China AF1627 FOXO1 Rabbit 1:1000 CST, USA 2880S Acetyl- FOXO1 Rabbit 1:1000 Affinity, USA AF2305 Cleaved caspase- 3 Rabbit 1:1000 CST, USA 9664S Bcl- 2 Rabbit 1:1000 ABclonal, China A0208 Bax Rabbit 1:1000 ABclonal, China A0207 β- Actin Rabbit 1:1000 Beyotime, China AF5003 T A B L E 1 The information of antibodies used.

Techniques: In Vitro, Western Blot, Expressing, Flow Cytometry

Journal: The FASEB Journal

Article Title: Ghrelin alleviates intestinal ischemia–reperfusion injury by activating the GHSR‐1α/Sirt1/FOXO1 pathway

doi: 10.1096/fj.202302155rrr

Figure Lengend Snippet: FIGURE 6 The ghrelin/GHSR-1α system promotes Sirt1 to protect against apoptosis. (A) Cell viability, (B) LDH release rate, (C) Western blotting analysis of GHSR-1α, Sirt1, ac-FOXO1, FOXO1, Bcl-2, Bax, and cleaved caspase3 expression, (D) quantification of the protein levels of GHSR-1α, Sirt1, ac-FOXO1/FOXO1, Bcl-2/Bax, and cleaved caspase3 in Caco-2 cells, (E) flow cytometry to detect the apoptosis rate, and (F) Statistical analysis of the apoptosis rate. Data are expressed as Mean ± SEM (n = 6). *p < .05.

Article Snippet: Antibodies Source Dilution Company Catalog number Ghrelin Rabbit 1:1000 Abmart, China TD6389S GHSR- 1α Rabbit 1:1000 Abmart, China TD2794S Sirt1 Rabbit 1:1000 CST, USA 9475S p- AMPK Rabbit 1:1000 CST, USA 50081S AMPK Rabbit 1:1000 Beyotime, China AF1627 FOXO1 Rabbit 1:1000 CST, USA 2880S Acetyl- FOXO1 Rabbit 1:1000 Affinity, USA AF2305 Cleaved caspase- 3 Rabbit 1:1000 CST, USA 9664S Bcl- 2 Rabbit 1:1000 ABclonal, China A0208 Bax Rabbit 1:1000 ABclonal, China A0207 β- Actin Rabbit 1:1000 Beyotime, China AF5003 T A B L E 1 The information of antibodies used.

Techniques: Western Blot, Expressing, Flow Cytometry

Journal: American Journal of Cancer Research

Article Title: PF-429242 exhibits anticancer activity in hepatocellular carcinoma cells via FOXO1-dependent autophagic cell death and IGFBP1-dependent anti-survival signaling

doi:

Figure Lengend Snippet: Role of FOXO1 in PF-429242-induced autophagic cell death in hepatocellular carcinoma (HCC) cells. (A) PLC5 and HepG2 cells were treated with 20 μM PF-429242 with or without indicated doses of AS1842856 (AS) for 24 h. Protein expressions were determined by Western blotting. (B) PLC5 and HepG2 cells were treated with 20 μM PF-429242 with or without 120 μM carbenoxolone (CBX) for 24 h. Protein expressions were determined by Western blotting. (C, D) PLC5 and HepG2 cells were transfected with siRNAs specific for FOXO1 in (C) or IGFBP1 (BP1) in (D) for 24 h, and then treated with 20 μM PF-429242 for 24 h. Protein expressions were determined by Western blotting. (E) si-FOXO1-transfected PLC5 and HepG2 cells were seeded in a 96-well plate and then treated with the indicated doses of PF-429242 for 72 h. Cell viability was determined by Alamar Blue staining. Error bars are the mean ± SD (n = 5). *** indicates a significant difference (P < 0.01) between si-FOXO1- and si-Control-transfected cells. (F) PLC5 and HepG2 cells were treated with 10, 20, and 40 μM PF-429242 for 24 h. Protein expressions were determined by Western blotting. Ratios between indicated protein expressions are shown.

Article Snippet: Acetyl-Lys262/Lys265/Lys274-FOXO1 (#A17406) and Ki67 (#A11390) antibodies were purchased from ABclonal Technology (Woburn, MA, USA).

Techniques: Western Blot, Transfection, Staining, Control