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

Journal: International Journal of Molecular Sciences

Article Title: Improved Wound Healing and Skin Regeneration Ability of 3,2′-Dihydroxyflavone-Treated Mesenchymal Stem Cell-Derived Extracellular Vesicles

doi: 10.3390/ijms24086964

Figure Lengend Snippet: List of antibodies for Western blot.

Article Snippet: Akt1 polyclonal antibody , rabbit , Cusabio (CSB-PA000855).

Techniques: Western Blot, Recombinant

Journal: Journal of Cellular and Molecular Medicine

Article Title: The preventive and therapeutic effects of AAV1‐KLF4‐shRNA in cigarette smoke‐induced pulmonary hypertension

doi: 10.1111/jcmm.16194

Figure Lengend Snippet: (A) Expression of KLF4 in the pulmonary vessels of rats in each group. Scale bar: 30 μm (B) Expression of PCNA in the pulmonary vascular smooth muscle of rats in each group. Scale bar: 25 μm (C) Representative bands of protein expression for KLF4 and PCNA in rat pulmonary vessels, and comparison of relative protein levels (normalized to GAPDH level). AU, arbitrary units. (D) Expression of P‐AKT in the pulmonary vessels of rats in each group. Scale bar: 20 μm. Sham (n = 8), saline (n = 6), AAV1‐control vector group (n = 6) and AAV1‐KLF4‐shRNA group (n = 11) at the end of the prevention experiment. Multiple comparisons were performed by one‐way ANOVA with SNK‐q. ** P < .01, *** P < .001

Article Snippet: Furthermore, the expression of KLF4 and P‐AKT was assessed by immunostaining with antibodies against KLF4 (ProteinTech, Wuhan, China) and P‐AKT (Gene Tex, San Antonio, USA).

Techniques: Expressing, Plasmid Preparation, shRNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: The preventive and therapeutic effects of AAV1‐KLF4‐shRNA in cigarette smoke‐induced pulmonary hypertension

doi: 10.1111/jcmm.16194

Figure Lengend Snippet: (A) Expression of KLF4 in the pulmonary vessels of rats in each group. Scale bar: 30 μm (B) Expression of PCNA in the pulmonary vascular smooth muscle of rats in each group. Scale bar: 25 μm (C) Representative bands of protein expression for KLF4 and PCNA in rat pulmonary vessels, and comparison of relative protein levels (normalized to GAPDH level). AU, arbitrary units. (D) Expression of P‐AKT in the pulmonary vessels of rats in each group. Scale bar: 20 μm. Sham (n = 6), saline (n = 7), AAV1‐control vector group (n = 7) and AAV1‐KLF4‐shRNA group (n = 8) at the end of the therapeutic experiment. Multiple comparisons were performed by one‐way ANOVA with SNK‐q. * P < .05, ** P < .01, *** P < .001

Article Snippet: Furthermore, the expression of KLF4 and P‐AKT was assessed by immunostaining with antibodies against KLF4 (ProteinTech, Wuhan, China) and P‐AKT (Gene Tex, San Antonio, USA).

Techniques: Expressing, Plasmid Preparation, shRNA

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