RNA was isolated 472 hr post-transfection and reversely transcribed using SuperScript VILO kit (Thermo Scientific). All qPCR analyses were performed with the QuantiTect Multiplex PCR kit (Qiagen, Hilden, Germany) on a StepOne Real-Time PCR Sytem (Applied Biosytems) with a total of 45 cycles. Constitutive maintenance gene 18S rRNA (Applied Biosystems, Quencher VIC/MGB, 4319413E) and human WRN (Applied Biosystems, Quencher FAM/MGB-NFQ, 4331182), human MLH1 (Applied Biosystems, Quencher FAM/MGB-NFQ, 4453320) and human MSH3 (Applied Biosystems, Quencher FAM/MGB-NFQ, 4448892) TaqMan probes were used. WRN expression was normalized to 18S rRNA expression levels and is indicated relative to the NTC control.
Lieb S., Blaha-Ostermann S., Kamper E., Rippka J., Schwarz C., Ehrenhöfer-Wölfer K., Schlattl A., Wernitznig A., Lipp J.J., Nagasaka K., van der Lelij P., Bader G., Koi M., Goel A., Neumüller R.A., Peters J.M., Kraut N., Pearson M.A., Petronczki M, & Wöhrle S. (2019). Werner syndrome helicase is a selective vulnerability of microsatellite instability-high tumor cells. eLife, 8, e43333.
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