GSM6979395: ATACseq_monocytes_AL_rep1; Mus musculus; ATAC-seq
Mice were euthanized with CO2, perfused via the left ventricle of the heart with 10 mL PBS before tissues were harvested and placed in cold PBS. Bone marrow was flushed using cold PBS and processed through a 70 mM filter. Spleens were processed through a 70 mM filter Whole bone marrow from femur or tibia was flushed in Trizol (Thermofisher). Total RNA extraction was performed with RNeasy mini kit (QIAGEN). DNase-treated total RNA (100 ng) was reverse transcribed into cDNA with Omniscript RT kit (QIAGEN) following manufacturer's instructions. qPCR was performed using the Taq PCR Master Mix (Life Technologies) on a Quantstudio 6 Flex Real-Time PCR (Life Technologies). CD8+ TCM were sorted from the bone marrow and spleen of mice fed ad libitum or on DR. RNA was extracted using an Arcturus PicoPure RNA Isolation Kit (Applied Biosystems), as per manufacturer's instructions. cDNA was synthesized with the Ultralow V4 kit (Clonetech) and sequencing libraries were subsequently prepared with the Nextera XT DNA prep kit (Illumina) and sequenced on NextSeq500.