Rna easy mini extract kit

Total mRNA was isolated from U251 cells by using RNA Easy Mini Extract Kit (Sigma-Aldrich, Gaithersburg, USA). Expressions of TGFβ, Fibronectin, Vimentin, E-cadherin, Bcl-2, Bcl-w, Caspase-3, and Bad in U251 cells were calculated using the RT-qPCR kit with β-actin expression as an endogenous control. All procedures were performed according to the manufacturer’s instructions. The forward and reverse primers were obtained from Invitrogen (Invitrogen, Shanghai, China) and β-actin levels were measured as an internal control. The expression level of mRNA was defined as mRNA/β-actin, measured by using the 2^−ΔΔCq protocol. PCR was repeated 3 times in triplicate.

Total mRNA was isolated from renal cells by using an RNA Easy Mini Extract kit (Sigma-Aldrich; Merck KGaA). RNA was reversed transcribed using a PrimeScript RT Master Mix kit (Takara Bio, Inc., Otsu, Japan). The expression of caspase-3, −8 and −9 as well as apoptotic protease activating factor 1 (Apaf-1) in renal cells was determined by using an RT-qPCR kit (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions with β-actin expression as an endogenous control. All procedures were performed according to the manufacturer's instructions. All of the primers were from Invitrogen (Thermo Fisher Scientific, Inc.) and their sequences are listed in Table I. Relative mRNA expression levels were determined by using the 2^−ΔΔCq method (31 (link)). The final results were presented as the fold of β-actin.