RNA-seq of LN activated B cells
Total RNA was extracted with the Arcturus PicoPure Kit and sequenced by GENEWIZ (Azenta) using poly(A) selection and ultra-low input library preparation. cDNA synthesis (SMART-Seq v4) and library preparation (Illumina Nextera XT) included fragmentation, adaptor addition, and limited-cycle PCR. Libraries were validated (Agilent TapeStation) and quantified (Qubit 3.0, qPCR). Sequencing (Illumina NovaSeq, 2150 bp PE) was followed by quality trimming (Trimmomatic v0.36), mapping to Mus musculus GRCm38 (STAR v2.5.2b), and FASTQ generation (bcl2fastq 2.20, one mismatch allowed)