rabbit anti p foxo1 s256  (Cell Signaling Technology Inc)

Journal: The Journal of Clinical Investigation

Article Title: Selective inhibition of long isoforms of phosphodiesterase 4D mitigates liver fibrosis in mouse models

doi: 10.1172/JCI182571

Figure Lengend Snippet: (A) PDGF-induced HSC proliferation (with or without D15987 ) was evaluated using BrdU incorporation assay. ( B ) LX-2 cells (left) and primary human HSCs (right) were treated with 10 ng/ml PDGF with or without D159687. Phosphorylated AKT was visualized using an antibody specific for ser473. ( C ) Phosphorylation of AKT in Kupffer cells. ( D ) Phosphorylation states of FOXO1 were determined by immunoblotting. ( E ) Confocal microscopy images of LX-2 cells stained with anti-FOXO1 (red) and DAPI (blue). ( F ) Expression of the cell cycle regulators. ( G ) Pde4d mRNA level in Kupffer cells stimulated with 1 μg LPS for 24 hours. ( H ) LX-2 Cell viability and caspase 3/7 activity under various concentrations of D159687 were evaluated. ( I ) Cleavage of PARP and caspase 3 in LX-2 cells upon D159687 treatment were evaluated by immunoblotting. ( J ) Cell viability was determined in LX-2 cells treated with D159687 in the presence of caspase inhibitor, Q-VD-Oph. ( K and L ) The effects of D159687 on the caspase 3/7 activity and viability of primary human hepatocytes. Staurosporine (STS, 2 μM) was used as a positive control of induction of apoptosis. All values are presented as the mean ± SEM of at least 3 independent experiments. 1-way ANOVA with Tukey’s post hoc test for multiple comparisons was used for statistical analysis in A , J , and K ; A 2-tailed unpaired Student’s t test was used to evaluate the statistical significance in G and L . ** P < 0.01; *** P < 0.001. Scale bar: 20 μl.

Article Snippet: Primary antibodies were used at 1:1,000 dilution, including COL1A1 (#84336, Cell signaling); ACTA2 (#A2547, Sigma-Aldrich); COL1A1 (#84336, Cell signaling); p-SMAD2 (#3108, Cell signaling); p-SMAD3 (#ab52903, abcam); SMAD2/3 (#8685, Cell signaling); SMAD4 (#46535, Cell signaling); LAMIN (#13435, Cell signaling); p-ERK (#4370, Cell signaling); ERK (#4696, Cell signaling); p-IKK (#2697, Cell signaling); IKK (#11930, Cell signaling); p-NF-κB p65 S536 (#3033, Cell signaling); NF-κB P65 (#8242, Cell signaling); p-AKT (#4060, Cell signaling); AKT (#4691, Cell signaling); p-FOXO1 S256 (#9461, Cell signaling); p-FOXO3a S253 (#13129, Cell signaling); FOXO1 (#2880, Cell signaling); p18 INK4C (#2896, Cell signaling); p21 Waf1/Cip1 (#2947, Cell signaling); CDK4 (#12790, Cell signaling); CDK6 (#3136, Cell signaling); PARP (#9542, Cell signaling); CASPASE3 (#14220, Cell signaling); Integrin a5 (#ab150361, abcam); p-FAK Y397 (#8556, Cell signaling); p-FAK Y925 (#3284, Cell signaling); FAK (#13009, Cell signaling); FLAG (#F1804, Sigma-Aldrich); a-tubulin (sc-8035, Santa Cruz).

Techniques: BrdU Incorporation Assay, Phospho-proteomics, Western Blot, Confocal Microscopy, Staining, Expressing, Activity Assay, Positive Control

Journal: Frontiers in Pharmacology

Article Title: Akkermansia muciniphila ameliorates olanzapine-induced metabolic dysfunction-associated steatotic liver disease via PGRMC1/SIRT1/FOXO1 signaling pathway

doi: 10.3389/fphar.2025.1550015

Figure Lengend Snippet: Effects of AKK and OLZ on the hepatic PGRMC1/SIRT1/FOXO1 signaling pathway. Relative mRNA expression of PGRMC1 (a) , SIRT1 (b) , FOXO1 (c) . Relative protein expression of PGRMC1 (d) , SIRT1 (e) , FOXO1 (f) , p-FOXO1 (g) . (h) The ratio of p-FOXO1/FOXO1. Differences were considered statistically significant when p < 0.05. * p < 0.05, ** p < 0.01, *** p < 0.001, HFD vs. LFD. # p < 0.05, ## p < 0.01, ### p < 0.001, OLZ vs. HFD. $ p < 0.05, $$ p < 0.01, $$$ p < 0.001, OLZ-AKK vs. OLZ-PBS.

Article Snippet: The antibodies used included PGRMC1 (1:5000; Proteintech; 12990-1-AP), SIRT1 (1:2000; HUABIO; ER130811), FOXO1 (1:1000; Proteintech; 18592-1-AP), p-FOXO1 (s256) (1:500; ProMab Biotechnologies Inc.; P40793), and β-actin (1:10000; Proteintech; 66009-1-Ig).

Techniques: Expressing