Revertra aceh kit

The amount of mRNA in the cultured HBMECs was measured using quantitative real-time PCR (qRT-PCR), according to standardized procedures. Twenty-four hours after the treatment, the total RNA was extracted from the cells with TRIzol reagent (Invitrogen, Carlsbad, CA, USA), following the manufacturer’s guidelines. After DNase treatment, the RNA was reverse transcribed to cDNA using the ReverTra AceH kit (Toyobo Co Ltd., Osaka, Japan) according to the manufacturer’s instructions. The cDNA underwent PCR using the Applied Biosystems 7500 Real-Time PCR System (Thermo Fisher Scientific, Waltham, MA, USA). The PCR primer sequences are shown in Table 1. The level of each gene was normalized to the β-actin values, and the reactions were performed in triplicate.

Total RNA was extracted from BMECs using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. After DNase treatment, RNA was reverse transcribed using the ReverTra AceH kit (Toyobo, Osaka, Japan) according to the manufacturer’s instructions. The cDNA was subjected to real-time PCR using an Applied Biosystems 7500 PCR System (Applied Biosystems, Foster City, CA, USA). The primer sequences and expected sizes of the PCR products are listed in Table 1. The expression levels for each gene of interest were normalized to their corresponding β-actin values. The reactions were run in triplicate.