Journal: Frontiers in Pharmacology

Article Title: Berberine Improves Chemo-Sensitivity to Cisplatin by Enhancing Cell Apoptosis and Repressing PI3K/AKT/mTOR Signaling Pathway in Gastric Cancer

doi: 10.3389/fphar.2020.616251

Figure Lengend Snippet: Berberine inhibited PI3K/AKT/mTOR signaling in the DDP-resistant gastric cancer cells with cisplatin treatment. (A,B) Western blot determined phosphor-PI3K, PI3K, phospho-AKT, AKT, phosphor-mTOR or mTOR protein levels in BGC-823/DDP and SGC-7901/DDP cells after treatment with DDP (30 µM), Berberine (30 µM) or DDP (30 µM) + Berberine (30 µM). * p < 0.05 compared to Blank group; # p < 0.05 compared to DDP (30 µM) group; $ p < 0.05 compared to berberine (30 µM) group ( n = 3).

Article Snippet: The concentrations and the sources of the primary antibodies were shown below: β-actin (1:2,000; Cell Signaling Technology, Danvers, United States), cleaved caspase-3 (1:1,000; Cell Signaling Technology), cleaved caspase-9 (1:1,000, Cell Signaling Technology), Bax (1:1,000; Cell Signaling Technology), multidrug resistance-associated protein 1 (MRP1; 1:1,000; Cell Signaling Technology), multi-drug resistance-1 (MDR1; 1:1,500; Cell Signaling Technology), phosphorylated (phospho)-PI3K (1:1,000; Cell Signaling Technology), PI3K (1:1,000; Cell Signaling Technology), phospho-AKT (1:1,000; Cell Signaling Technology), AKT (1:1,000; Cell Signaling Technology), phospho-mTOR (1:1,000; Cell Signaling Technology) and mTOR (1:1,000; Cell Signaling Technology).

Techniques: Western Blot

Journal: Molecular Medicine Reports

Article Title: Penehyclidine hydrochloride inhibits TLR4 signaling and inflammation, and attenuates blunt chest trauma and hemorrhagic shock-induced acute lung injury in rats

doi: 10.3892/mmr.2018.8644

Figure Lengend Snippet: PHC treatment inhibits the THS-induced expression of TLR4 and p-p38MAPK. (A) Western blot analysis revealed that the expression of (B) TLR4 and (C) p-p38MAPK in the THS group was increased compared with that in the sham group. PHC attenuated the increase in the expression of TLR4 and p-p38MAPK compared with that in the THS group. The expression of p-p38MAPK in the lung tissue was examined under a light microscope following (D) the immunofluorescence assay (magnification, ×200), and (E) the expression was quantified. *P<0.05 vs. sham group; # P<0.05 vs. THS group. The data are presented as the mean ± standard deviation (n=10). THS, blunt chest trauma with hemorrhagic shock; PHC, penehyclidine hydrochloride; p, phosphorylated; MAPK, mitogen-activated protein kinase; TLR4, Toll-like receptor 4.

Article Snippet: TLR4 and phosphorylated (p)-p38MAPK antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

Techniques: Expressing, Western Blot, Light Microscopy, Immunofluorescence, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: Baicalin and baicalein attenuate renal fibrosis in vitro via inhibition of the TGF-β1 signaling pathway

doi: 10.3892/etm.2017.4888

Figure Lengend Snippet: Baicalin and baicalein downregulate TGF-β1-induced endogenous TGF-β1 expression and SMAD3 activation. Cells cultured in COLI-coated plates were treated with 20–80 µM (A) baicalin or (B) baicalein. After 48 h, cells were collected for RT-qPCR analysis of TGF-β1 mRNA expression. Cells were treated with 80 µM (C) baicalin or (D) baicalein and collected at 24, 48, 72 and 96 h for RT-qPCR analysis of TGF-β1 mRNA expression. (E) After 48 h treatment with 20–80 µM baicalin or baicalein double immunofluorescence staining was performed to visualize the expression of α-SMA (red) and phosphorylated-SMAD3 (green). HEK-293T cells were transfected with the pSMAD-binding element-SEAP plasmid and then treated with 20–80 µM (F) baicalin or (G) baicalein with TGF-β1. After 48 h treatment, supernatant was collected and analyzed for SEAP expression. **P<0.01, ***P<0.001 vs. TGF-β1 treatment alone. TGF-β1, transforming growth factor β1; SMAD3, mothers against decapentaplegic homolog 3; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; SEAP, secreted embryonic alkaline phosphatase; COLA1, collagen type I α1; α-SMA, α smooth muscle actin.

Article Snippet: The SEAP signal was quantified using the SensoLyte ® p-nitrophenyl phosphate (pNPP) SEAP Reporter Gene assay kit (AnaSpec, Fremont, CA, USA) according to the manufacturer's protocol.

Techniques: Expressing, Activation Assay, Cell Culture, Quantitative RT-PCR, Double Immunofluorescence Staining, Transfection, Binding Assay, Plasmid Preparation, Reverse Transcription, Real-time Polymerase Chain Reaction