Journal: Biochemistry and Biophysics Reports

Article Title: Effects of branched-chain amino acids on iron deficiency-induced muscle atrophy

doi: 10.1016/j.bbrep.2026.102451

Figure Lengend Snippet: BCAA increased the p-Akt in DFO-treated myotubes after 45 min of BCAA treatment, but had no statistically significant effects on other signaling molecules at the same time point. Representative blots for quantification of (A) p-Akt (F (3, 8) = 6.166, p = 0.018, η 2 = 0.6981), p-mTOR, p-p70S6K, p-4E-BP1, and p-eEF2, (B) p-AMPK, and p-ACC, and (C) p-FOXO1, and p–NF–κB p65. Protein was extracted at 45 min after BCCA treatment. Data are presented as the mean ± SD (error bars) from three independent experiments (N = 3), and were analyzed using one-way analysis of variance (ANOVA) with eta squared (η 2 ) used to measure effect sizes, followed by a post hoc Tukey–Kramer test. ∗p < 0.05.

Article Snippet: p-FoxO1 (Ser256) , Rabbit , 84192 , 1:1000 , Cell Signaling Technology.

Techniques:

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: Experimental and Therapeutic Medicine

Article Title: Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway in vitro

doi: 10.3892/etm.2025.12956

Figure Lengend Snippet: Role of the FoxO signaling pathway in pc-SLC2A4-induced changes in HG-induced HTR-8/SVneo cells. (A) Screen of targeted pathways related to gestational diabetes mellitus and SLC2A4 (five common genes identified through the CTD/GeneCards/DisGeNet database) using Kyoto Encyclopedia of Genes and Genomes, suggesting a critical role for FoxO signaling pathway. (B) The expression levels of p-FoxO1, FoxO1, p-FoxO3a and FoxO3a were analyzed by western blotting, GAPDH was used as the internal control for the results. The expression of INS and INSR was analyzed by (C) reverse transcription-quantitative PCR and (D) western blotting. (E) Glucose uptake was detected by glucose uptake assay. (F) Cell viability was measured using a Cell Counting Kit-8 assay. *** P<0.001 vs. Control; ns P>0.05 vs. HG; ### P<0.001 vs. HG + pc-NC; $ P<0.05, $$ P<0.01 and $$$ P<0.001 vs. HG + pc-SLC2A4. SLC2A4, solute carrier family 2 member 4; HG, high glucose; ns, not significant; NC, negative control; OD, optical density; INS, insulin; INSR, INS receptor; p, phosphorylated; FoxO, Forkhead box O.

Article Snippet: The membranes were then incubated overnight at 4 ̊C with primary antibodies against SLC2A4 (cat. no. 2213S; Cell Signaling Technology, Inc.), INSR (cat. no. 3025S; Cell Signaling Technology, Inc.), INS (cat. no. ab181547; Abcam), phosphorylated (p)-FoxO1 (cat. no. 9461S; Cell Signaling Technology, Inc.), FoxO1 (cat. no. 2880S; Cell Signaling Technology, Inc.), p-FoxO3a (cat. no. 13129S; Cell Signaling Technology, Inc.), FoxO3a (cat. no. 12829S; Cell Signaling Technology, Inc.) and GAPDH (cat. no. 97166; Cell Signaling Technology, Inc.) (all 1:1,000 dilution).

Techniques: Expressing, Western Blot, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Counting, Negative Control