Journal: Molecular medicine reports

Article Title: Expression of key enzymes in the mevalonate pathway are altered in monocrotaline-induced pulmonary arterial hypertension in rats.

doi: 10.3892/mmr.2017.7798

Figure Lengend Snippet: Figure 2. Expression levels of HMGR, FDPS, FNTA, FNTB and GGTase-I in the MCT-induced PAH rat pulmonary artery. The proteins were extracted from pulmonary arteries of each group. (A) Western blot analyses demonstrate the expression of HMGR, FDPS, FNTA, FNTB and GGTase-I in the control and MCT-induced PAH rats. GAPDH was used as the endogenous loading control. Graphs demonstrate the relative changes in (B) HMGR, (C) FDPS, (D) FNTA, (E) FNTB and (F) GGTase-I in the control and MCT groups. Data are expressed as the mean ± standard deviation. *P<0.05 and **P<0.01 compared with the control group. PAH, pulmonary arterial hypertension; MCT, monocrotaline; HMGR, 3-hydroxy-3-methylglutaryl-coenzyme A; FDPS, farnesyldiphosphate synthase; FNTA, farnesyltransferase α; FNTB, farnesyltransferase β; GGTase-I, geranylgeranyltransferase type I.

Article Snippet: The membrane was blocked in 5% skim milk (5 g skim milk dissolved in 100 ml Tris-buffered saline Tween solution) at room temperature for 1 h. The membrane was incubated with the following antibodies: HMGR (cat. no. ab174830, 1:2,000), FDPS (cat. no. ab189874, 1:1,000), FNTA (cat. no. ab109738, 1:1,000), and FNTB (cat. no. ab109748, 1:1,000) (all from Abcam, Cambridge, UK), GGT-I (cat. no. sc18996, 1:200; Santa Cruz Biotechnology Co., Ltd., Dallas, TX, USA), phosphorylated (p)-eNOS (cat. no. 95719, 1:1,000), eNOS (cat. no. 9586, 1:1,000), and RhoA (cat. no. ARH04, 1:1,000) (all from CST Biological Reagents Company Limited, Shanghai, China), Rac1 (cat. no. ARC03, 1:1,000; BD Biosciences, Franklin Lakes, NJ, USA) at 4 ̊C for 16 h. The membranes were then incubated with the appropriate secondary antibody: Goat-anti-rabbit immunoglobulin G (IgG) (cat. no. 1268, 1:2,500), goat-anti-mouse IgG (cat. no. 1265, 1:2,500), and rabbit-anti-goat IgG (cat. no. M1102, 1:2,500) (all from Biovision, Inc., Milpitas, CA, USA) for 2 h at room temperature.

Techniques: Expressing, Western Blot, Control, Standard Deviation