RNA Extraction and qPCR Analyses of PBMCs

Total RNA was purified from freshly thawed (U)PBMC using the Qiagen RNeasy Kit (QIAGEN, Hilden, Germany) according to the manufacturer’s instructions. The RNA yield was quantified with the Qubit RNA broad range (BR) assay in combination with a Qubit spectrophotometer (Life Technologies). Samples were stored in RNase free water (Life Technologies) at −20 °C for a maximum of 7 days prior to cDNA transcription (see SI, section for further details). As previously reported [9 (link)] we selected Succinate dehydrogenase complex, subunit A, flavoprotein variant (SDHA) and Importin 8 (IPO8) as gene expression references. Detailed information about the gene expression analyses and the selection procedure of appropriate house-keeping genes for human (U)PBMC is given in the SI, section 4.