Journal: Drug Design, Development and Therapy

Article Title: Eucommia bark (Du-Zhong) improves diabetic nephropathy without altering blood glucose in type 1-like diabetic rats

doi: 10.2147/DDDT.S98558

Figure Lengend Snippet: Effect of Du-Zhong on phosphorylated Smad 2/3 level in renal tissues of rats. Notes: ( A ) Effects of treatments on p-Smad2/3 in renal tissues of rats. ( B ) STZ-diabetic rats were dosed by oral gavage once daily for 20 days with 1 g/kg Du-Zhong (STZ + DZ). Normal or STZ-diabetic rats received the vehicle treatment at the same volume of vehicle used to prepare the test solution. Ratio of p-Smad2/3/actin is expressed as the mean ± standard deviation (n=4 per group) in each column. *** P <0.001 compared with data obtained from vehicle-treated normal Wistar rats (Wistar). ## P <0.01 compared with data obtained from vehicle-treated STZ-diabetic rats (STZ). Abbreviations: DZ, Du-Zhong; p-Smad2/3, phosphorylated Smad2/3; STZ, streptozotocin.

Article Snippet: Membranes were blocked with 5% nonfat dry milk in Tris-buffered saline Tween (20 mmol/L Tris, pH 7.6, 137 mmol/L NaCl, and 0.1% Tween 20) for 3 hours at room temperature, followed by an overnight incubation at 4°C with polyclonal antibodies against rat transforming growth factor-beta 1 (TGF-β1) (Abcam plc, Cambridge, UK; Cat No ab66043), connective tissue growth factor (CTGF; Abcam plc; Cat No ab6992), signal transducers and activators of transcription 3 (STAT3; Abcam plc; Cat No ab50761), phosphorylated (p-Smad2/3) antibody (Ser 423/425) (Santa Cruz Biotechnology Inc., Dallas, TX, USA; Cat No sc-11769), or actin (Santa Cruz Biotechnology Inc.; Cat No sc-1616).

Techniques: Standard Deviation

Journal: Oncology Letters

Article Title: TGFBR1*6A is a potential modifier of migration and invasion in colorectal cancer cells

doi: 10.3892/ol.2018.7725

Figure Lengend Snippet: To investigate the potential mechanisms underlying the TGFBR1*6A-induced switch from TGF-β1-mediated growth inhibition to growth stimulation in SW48 colorectal cancer cells, western blotting was performed. Western blotting demonstrating that increased expression of p-p38 and p-ERK1/2 were detected within 15–30 min of stimulation with exogenous TGF-β1 (5 ng/ml) in SW48 cells transfected with a *6A plasmid. When treated with TGF-β1 (5 ng/ml), the wild type SW48 (TGFBR1*9A) and controls cells activated p-Smad2 signaling, but only induced little activation of p-p38 and p-ERK signaling. However, under the same conditions in SW48-*6A cells, TGF-β1 activated both p-p38 and p-ERK signaling, while the expression of p-Smad2 was decreased. TGFBR1*6A, type 1 transforming growth factor β receptor; TGF-β1, transforming growth factor-β1; ERK, extracellular-signal-regulated kinase; MAPK, mitogen-activated protein kinase; p-, phosphorylated; Smad2, SMAD family member 2.

Article Snippet: Subsequently, the membranes were incubated at 4°C overnight with the following antibodies: Mouse monoclonal antibodies against β-actin (dilution, 1:1,000; cat. no. sc-70319; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and SMAD family member 2 (Smad2; dilution, 1:500; cat. no. sc133098; Santa Cruz Biotechnology, Inc.), and rabbit polyclonal antibodies against phosphorylated (p)-Smad2 (Ser465/467; dilution, 1:800; cat. no. 8828; Cell Signaling Technology, Inc., Danvers, MA, USA), p38 MAPK (dilution, 1:800; cat. no. 8690; Cell Signaling Technology, Inc.), p-p38 (Thr180/Tyr182) MAPK (dilution, 1:1,000; cat. no. 9211; Cell Signaling Technology, Inc.), extracellular-signal-regulated kinases 1/2 (ERK1/2; dilution, 1:800; cat. no. 9102; Cell Signaling Technology, Inc.) and p-Erk1/2 (Thr202/Tyr204; dilution, 1:800; cat. no. 9106; Cell Signaling Technology, Inc.).

Techniques: Inhibition, Western Blot, Expressing, Transfection, Plasmid Preparation, Activation Assay