phosphorylated p foxo1  (Cell Signaling Technology Inc)

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

Journal: Preventive Nutrition and Food Science

Article Title: Standardized Fingerroot ( Boesenbergia pandurata ) Extract Decelerates the Development of Sarcopenia in Aged Rats

doi: 10.3746/pnf.2025.30.1.47

Figure Lengend Snippet: Effects of Boesenbergia pandurata extract (BPE) on protein-degradation-related factors. (A) MuRF1 and atrogin-1 mRNA expression. Values were normalized to the expression of β-actin. (B) p-FoxO3a and FoxO3a protein expression levels. Values were normalized to the expression of the total form of FoxO3a. β-Actin and α-tubulin were used as loading controls. Total RNA and protein were isolated from gastrocnemius muscle tissues. Values are presented as the mean±SD (n=4 per group). # P <0.05, ## P <0.01 (vs. Young group); * P <0.05, ** P <0.01 (vs. Aged group).

Article Snippet: After washing with in Tris-buffered saline containing Tween 20 (TBST), the membranes were incubated with primary antibodies against FoxO3a, phosphorylated (p)-FoxO3a Thr32, PI3K, p-PI3K Tyr458, Akt, p-Akt Ser473, mTOR, p-mTOR Ser2448, p70S6K, p-p70S6K Thr389, 4EBP1, p-4EBP1 Thr37/46, and α-tubulin (1:1,000 dilution; Cell Signaling) overnight at 4°C.

Techniques: Expressing, Isolation

Journal: International Journal of Biological Sciences

Article Title: CDH1 is Identified as A Therapeutic Target for Skin Regeneration after Mechanical Loading

doi: 10.7150/ijbs.51309

Figure Lengend Snippet: E-cadherin knockdown induces keratinocyte dedifferentiation through the β-catenin-FOXO1-KLF4 pathway. ( A ) E-cadherin knockdown promoted β-catenin and FOXO1 nuclear localization. Image shows immunofluorescence staining of β-catenin (red) and FOXO1 (green). ( B ) Western blot of FOXO1 nuclear localization signal after E-cadherin knockdown. ( C ) Western blot of FOXO1 dephosphorylation state after E-cadherin knockdown. ( D ) Immunoprecipitation demonstrating β-catenin and FOXO1 interaction after E-cadherin knockdown. ( E ) FOXO1 ChIP-PCR showing significantly higher relative DNA levels of KLF4, KRT5, and KRT15 after E-cadherin knockdown as compared to the control group. N = 3 experiments. * P < 0.05, ** P < 0.01.

Article Snippet: The primary antibodies used were antibodies against E-cadherin (1:200; BD Biosciences), KRT5 (1:200; Abcam), KRT15 (1:100; Abcam), KRT19 (1:200; Abcam), phosphorylated (p)-FOXO1 (1:1000; Cell Signaling Technology, Beverly, MA, USA), FOXO1 (1:1000; Cell Signaling Technology), β-catenin (1:1000; Cell Signaling Technology), β-actin (1:1000; Cell Signaling Technology), and histone (1:500; Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Staining, Western Blot, De-Phosphorylation Assay, Immunoprecipitation

Journal: International Journal of Biological Sciences

Article Title: CDH1 is Identified as A Therapeutic Target for Skin Regeneration after Mechanical Loading

doi: 10.7150/ijbs.51309

Figure Lengend Snippet: Effect of KLF4 and FOXO1 on stemness markers in human keratinocytes. ( A ) Immunofluorescence staining of KLF4 and FOXO1 showing that E-cadherin knockdown promoted their nuclear localization. ( B ) Immunofluorescence staining of KLF4 and FOXO1 in human skin tissue before and after mechanical loading. Stretched skin showed significantly more KLF4 and FOXO1 nuclear localization than normal skin. ( C ) KLF4 ChIP-PCR indicating significantly higher relative DNA levels of KRT14, KRT19, and KRT15 after E-cadherin knockdown as compared to the control group. N = 3 experiments. ** P < 0.01. ( D ) qPCR of siKLF4 keratinocytes: stemness markers were repressed after interference. N = 3 experiments. ** P < 0.01. ( E ) qPCR of siFOXO1 keratinocytes: stemness markers were repressed after interference. N = 3 experiments. ** P < 0.01. ( F ) qPCR of CDH1/KLF4 or CDH1/FOXO1 keratinocytes: stemness markers were repressed after interference; CDH1/KLF4 or CDH1/FOXO1 double interference diminished the dedifferentiation caused by CDH1 knockdown. N = 3 experiments.

Article Snippet: The primary antibodies used were antibodies against E-cadherin (1:200; BD Biosciences), KRT5 (1:200; Abcam), KRT15 (1:100; Abcam), KRT19 (1:200; Abcam), phosphorylated (p)-FOXO1 (1:1000; Cell Signaling Technology, Beverly, MA, USA), FOXO1 (1:1000; Cell Signaling Technology), β-catenin (1:1000; Cell Signaling Technology), β-actin (1:1000; Cell Signaling Technology), and histone (1:500; Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Staining