Journal: BMJ Open Diabetes Research & Care

Article Title: Mouse model of metformin-induced diarrhea

doi: 10.1136/bmjdrc-2019-000898

Figure Lengend Snippet: Effects of wood creosote on diabetic obese mice treated with metformin. Wood creosote does not affect efficacy of metformin. Weight change (A) and blood glucose levels (fed condition) (B) were examined during the experiment in . Data are shown as means±SEM. No significant differences between the metformin group and the metformin with wood creosote (M+C) group were observed. Mice were fasted for 3 hours after the final administration of drugs. (C) One g/kg glucose was administrated via oral gavage. One to 2 µL blood was collected via tail vein at indicated time points, and blood glucose was measured by using GlucoSensor. (D) At day 4 of , mice were fasted for 2 hours and then injected intraperitoneally with 36 µg/kg insulin. Blood glucose levels are shown as % initial. At day 5 in , blood was collected and measured serum bile acid levels (E) and alanine aminotransferase (F) were measured. (G) Western blots were carried out using liver protein (50 µg) to detect levels of phospho-AMPK and phoshopho-AKT. AKT, Protein Kinase B; AMPK, AMP-activated kinase.

Article Snippet: Antibodies are as follows: Anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1E6D9, Proteintech, Rosemont, Illinois, USA), anti-AMPK (ABV10739, ABGENT, San Diego, California, USA), anti-phospho AMPK (pT172) (40H9, Cell Signaling Technology, Danvers, Massachusetts, USA), anti-total Protein Kinase B (AKT) (200–401 N98, Rockland, Limerick, Pennsylvania, USA), and anti-phosho AKT (pS473) (D9E, Cell Signaling Technology).

Techniques: Injection, Western Blot

Journal: Scientific Reports

Article Title: LRP12 promotes gastric cancer progression through AKT/mTOR signaling and M2 macrophage-mediated immunosuppression

doi: 10.1038/s41598-025-28503-7

Figure Lengend Snippet: LRP12 enhances gastric cancer progression via the AKT/mTOR pathway. A-B Enrichment of the PI3K-AKT-mTOR signaling pathway in the STAD- GSE167297 cohort. C-D The protein levels of the AKT/mTOR signaling pathway in AGS cells with the control and stably overexpressing LRP12 were determined by Western Blot. All lanes shown in this blot image were derived from the same gel and membrane. Western blotting experiments were repeated in three independent biological. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Proteins (30 μg) were separated by SDS-PAGE, transferred to PVDF membranes (Millipore, USA), blocked with 5% skimmed milk for 2 h at room temperature, and incubated overnight at 4 °C with primary antibodies: LRP12 (ab150352, Abcam, UK, 1:2000), Tubulin (66031-1-Ig, ProteinTech, China, 1:5000), phospho-mTOR (Ser2448) (80596-1-RR, ProteinTech, 1:2000), mTOR (66888-1-Ig, ProteinTech, 1:2000), phospho-AKT (Ser473) (80455-1-RR, ProteinTech, 1:2000), and AKT (10176-2-AP, ProteinTech, 1:2000).

Techniques: Control, Stable Transfection, Western Blot, Derivative Assay, Membrane

Journal: Hepatology (Baltimore, Md.)

Article Title: The chemosensory function of primary cilia regulates cholangiocyte migration, invasion and tumor growth

doi: 10.1002/hep.30308

Figure Lengend Snippet: (A,B) Western blots showing the effect of ATP for 30 minutes on AKT and PTEN phosphorylation in normal ciliated cholangiocytes (NHC SCR), experimentally deciliated cholangiocytes (NHC IFT88), and the iCCA cell line HUCTT1. Bar graph shows densitometry expressed as % of phosphorylated/total ratios in control conditions (*p<0.05, n=3) (**p<0.01, n=3).

Article Snippet: After blocking, the membranes were incubated with the appropriate primary antibodies against IFT88 (ProSci, Poway, CA), LKB1, p-LKB1(S428), AKT, p-AKT(S473), PTEN and p-PTEN(Ser380/Thr382/Thr383) (Cell Signaling), FAK (Cell Signaling) and GAPDH (ProSci) at 4°C overnight.

Techniques: Western Blot

Journal: Hepatology (Baltimore, Md.)

Article Title: The chemosensory function of primary cilia regulates cholangiocyte migration, invasion and tumor growth

doi: 10.1002/hep.30308

Figure Lengend Snippet: Nucleotides are detected by P2Y11 receptor localized in the primary cilium activating AC5, which induces increases in the levels of cAMP and activation of PKA. Once activated PKA phosphorylates and activates LKB1, LKB1 phosphorylates and stabilizes PTEN, leading to AKT inhibition. Inhibited AKT is unable to phosphorylate F-actin that is required to reorganize the cytoplasm and protrusions in migrating cells. HMC can activate LKB1 via a ciliary-independent mechanism emulating the chemosensory function of primary cilia.

Article Snippet: After blocking, the membranes were incubated with the appropriate primary antibodies against IFT88 (ProSci, Poway, CA), LKB1, p-LKB1(S428), AKT, p-AKT(S473), PTEN and p-PTEN(Ser380/Thr382/Thr383) (Cell Signaling), FAK (Cell Signaling) and GAPDH (ProSci) at 4°C overnight.

Techniques: Activation Assay, Inhibition