Cyber green pcr master mix

Total RNA was isolated from the lung lobe, liver, and gut of Rosa Vav1 mice after Vav1 transgene induction, using TRI reagent (Sigma Kiryat HaMada, Jerusalem, Israel), and the miRNeasy Kit (QIAGEN, Maryland, MD, USA) according to the manufacturer’s instructions. cDNAs were prepared using the qScript reverse transcriptase (Quanta, Taipei, Taiwan). Quantitative PCR (qPCR) was performed by using the CFX-384 real-time PCR machine (Bio-Rad, Hercules, CA, USA) to detect Vav1 transgene, mVAV1, and GFP using cyber green PCR master mix (Bio-Rad, Hercules, CA, USA) and the required primers (Supplementary Table S1B). Three independent experiments were performed, and in each experiment, triplicates were used.

Total RNA and cDNAs were prepared as detailed for reverse transcriptase-PCR. Detection of VAV1, SMA, and MMP2 was performed using cyber green PCR master mix (BioRad, CA, USA) and the required primers (Table S1). Real-time PCR was performed by using the ABI Prism 7300 real-time PCR technology (Applied Biosystems, CA, USA). Three independent experiments were performed, and in each experiment triplicates were used.