Bulk rna seq of ccr2ko male mus musculus
RNA was extracted using the RNeasy mini kit (Qiagen) according to the manufacturers instructions. Quality control and sequencing was done by Novogene CO., Ltd. (Sacramento, CA). Briefly, RNA integrity and concentration were assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies) and a nanodrop. Only samples with RNA10 integrity number (RIN) > 7 were used for library preparation. mRNA was enriched using poly-T oligo-attached magnetic beads, fragmented, and reverse transcribed using random hexamer primers. RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep kit for Illumina (New England Biolabs), following the manufacturers protocol. Library quality was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Sequencing was performed on the Illumina NovaSeq 6000 platform, yielding paired-end reads with an average depth of approximately 25 million reads per sample.