Journal: Gastroenterology

Article Title: Neurotensin signaling activates microRNAs-21 and -155 and Akt, promotes tumor growth in mice, and is increased in human colon tumors.

doi: 10.1053/j.gastro.2011.07.038

Figure Lengend Snippet: Figure 4. NT regulates miR-21 and miR-155 signaling pathways in colon cancer cells. (A) Luciferase activity of the PTEN and SOCS1 3=UTRs after NT treatment (100 nmol/L) for 24 hours in HCT116 and DLD1 cells (untreated, as-miR-NC–treated, as-miR-21–treated, or as-miR-155–treated). (B) PTEN and SOCS1 mRNA levels, assessed by real-time PCR, in NT-treated (100 nmol/L, 6 h) HCT116 and DLD1 cells (untreated, as-miR-NC– treated, as-miR-21–treated, or as-miR-155–treated). (C) PTEN, SOCS1, and -actin protein levels, assessed by Western blot, in NT-treated (100 nmol/L, 6 h) HCT116 and DLD1 cells. (D) AKT phosphorylation (S473) levels, assessed by enzyme-linked immunosorbent assay, in HCT116 and DLD1 cells transfected with as-miR-NC (100 nmol/L), as-miR-21 (100 nmol/L), and/or as-miR-155 (100 nmol/L) for 24 hours and treated with NT (100 nmol/L) for 24 hours.

Article Snippet: AKT phosphorylation (S473) (DuoSet IC nzyme Linked Immunosorbent Assay, DYC887; R&D Systems, inneapolis, MN), phospho-GSK3 (S9) (DYC-1590; R&D Sysems), and phospho-p65(S536) (7834; Cell Signaling) were asessed by enzyme linked immunosorbent assay in cells transected with as-miR-21 (100 nmol/L) and/or as-miR-155 (100 mol/L) for 24 hours and treated with NT (100 nmol/L) for 24 ours.

Techniques: Protein-Protein interactions, Luciferase, Activity Assay, Real-time Polymerase Chain Reaction, Western Blot, Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Transfection

Journal: Gastroenterology

Article Title: Neurotensin signaling activates microRNAs-21 and -155 and Akt, promotes tumor growth in mice, and is increased in human colon tumors.

doi: 10.1053/j.gastro.2011.07.038

Figure Lengend Snippet: Figure 5. NT activates AKT through suppression of PPP2CA by direct interaction with miR-155. (A) miR-155 binding sites in 3=UTRs of PPP2CA predicted by Lever algorithm analysis. (B) Luciferase activity of the PPP2CA 3=UTRs in HCT116 and DLD1 cells transfected with as-miR-155 (100 nmol/L) or as-miR-155 (100 nmol/L) for 24 hours. (C) Luciferase activity of the PPP2CA 3=UTRs in HCT116 and DLD1 cells treated with NT (100 nmol/L). (D) PPP2CA and -actin protein levels, assessed by Western blot, in NT-treated (100 nmol/L, 6 h) HCT116 and DLD1 cells. (E) AKT phosphorylation (S473) levels, assessed by enzyme-linked immunosorbent assay, in HCT116 and DLD1 cells transfected with miR-155 (100 nmol/L) or si-PPP2CA (100 nmol/L) for 24 hours. (F) NF-B/p65 activity, assessed by enzyme-linked immunosorbent assay, in HCT cells transfected with NT (100 nmol/L) and as-miR-NC (100 nmol/L), as-miR-21 (100 nmol/L), as-miR-155 (100 nmol/L), or with 10 nmol/L of an AKT pharmacologic inhibitor (MK-2206).

Article Snippet: AKT phosphorylation (S473) (DuoSet IC nzyme Linked Immunosorbent Assay, DYC887; R&D Systems, inneapolis, MN), phospho-GSK3 (S9) (DYC-1590; R&D Sysems), and phospho-p65(S536) (7834; Cell Signaling) were asessed by enzyme linked immunosorbent assay in cells transected with as-miR-21 (100 nmol/L) and/or as-miR-155 (100 mol/L) for 24 hours and treated with NT (100 nmol/L) for 24 ours.

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Western Blot, Phospho-proteomics, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Interferon-γ Suppresses Intestinal Epithelial Aquaporin-1 Expression via Janus Kinase and STAT3 Activation

doi: 10.1371/journal.pone.0118713

Figure Lengend Snippet:

Article Snippet: Phosphorylated AKT (Cat# AF887, R & D Systems, Minneapolis, MN) , 5% BSA in TTBS , 1:400 , goat anti-rabbit.

Techniques: Blocking Assay, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Resistance training alleviates muscle atrophy and muscle dysfunction by reducing inflammation and regulating compromised autophagy in aged skeletal muscle

doi: 10.3389/fimmu.2025.1597222

Figure Lengend Snippet: Resistance training regulated mTORC1-HIF-1α pathway and alleviated chronic inflammation. (A) Representative Western blot images of mTOR, Raptor, HIF-1α, p70S6K, p-AKT (Ser473), AKT, IL-1β, and the internal control GAPDH in the quadriceps muscle. (B–H) Protein expression levels of p-AKT (Ser473), p-AKT (Ser473)/AKT, mTOR, Raptor, HIF-1α, p70S6K, and IL-1β. All data are presented as means ± SEM (two-way ANOVA with Tukey post hoc test), n = 6 per group; *p < 0.05, **p < 0.01.

Article Snippet: Membranes were blocked at room temperature for 1 h with 5% non-fat dry milk (Bio-Rad, 1706404) in Tris-buffered saline with Tween-20 (TBST) and incubated overnight at 4°C with primary antibodies: p-AKT (Ser473) (Cell Signaling Technology, #86758, 1:1000), AKT (Cell Signaling Technology, #9272, 1:1000), p-AMPKα (Thr172) (Cell Signaling Technology, #2535, 1:1000), AMPKα (Cell Signaling Technology, #2532, 1:1000), mTOR (Cell Signaling Technology, #2983, 1:1000), Raptor (Cell Signaling Technology, #2280, 1:1000), p70S6K (Cell Signaling Technology, #9202, 1:1000), IL-1β (Proteintech, 16806-1-AP, 1:2000), HIF-1α (Cell Signaling Technology, #14179, 1:1000), Beclin1 (Proteintech, 11306-1-AP, 1:1000), TFEB (Proteintech, 13372-1-AP, 1:1000), p62 (Proteintech, 18420-1-AP, 1:1000), LC3 (Cell Signaling Technology, #2775, 1:2000), and GAPDH (Proteintech, 60004-1-Ig).

Techniques: Western Blot, Control, Expressing

Journal: Molecular medicine reports

Article Title: Chinese herb extract improves liver steatosis by promoting the expression of high molecular weight adiponectin in NAFLD rats.

doi: 10.3892/mmr.2017.7284

Figure Lengend Snippet: Figure 5. Expression of proteins in adipose tissue analyzed using western blot analysis. Results of the western blot analysis are showed in graph. (A and B) Optical density value ratios of target protein to GAPDH are showed in bar graphs. **P<0.01, T group vs. M group; §§P<0.01, T group vs. CK group; #P<0.05, M group vs. CK group; ##P<0.01, M group vs. CK group. CK, normal control group; M, model control group; T, treatment group; HMW APN, high molecular weight adiponectin; FOXO1, forkhead box O protein 1; DsbA‑L, disulfide‑bond A oxidoreductase‑like protein; p‑, phosphorylated.

Article Snippet: The primary antibodies were as follows: GADPH (cat. no. AP0063; 1:5,000; Bioworld Technologies, Inc., St. Louis Park, MN, USA), Akt (cat. no. ab9150; 1:2,000; Abcam, Cambridge, UK), phosphorylated (p)-Akt (cat. no. 2965; 1:1,000; Cell Signaling Technology, Inc., Danvers, MA USA), FOXO1 (cat. no. 2880; 1:10,00; Cell Signaling Technology, Inc.), p-FOXO1 (cat. no. 9461; 1:1,000; Cell Signaling Technology, Inc.), DsbA-L (cat. no. ab92819; 1:2,000; Abcam, Cambridge, UK) and APN (cat. no. bs-0471R; 1:300; Bioss, Beijing, China).

Techniques: Expressing, Western Blot, Control, High Molecular Weight

Journal: Frontiers in Oncology

Article Title: mTOR Inhibitors as Radiosensitizers in Neuroendocrine Neoplasms

doi: 10.3389/fonc.2020.578380

Figure Lengend Snippet:

Article Snippet: Phospho-Akt (Ser473) (D9W9U) , Mouse, monoclonal , 1:1000 in 5% BSA/TBST , #12694, Cell Signaling (Danvers, US).

Techniques: