GSM9451574: GFP negative Tfh cells 4; Mus musculus; RNA-Seq
Spleens from SRBC-injected ChAT-GFP-tg animals were harvested on day 8 after immunization. ChAT-GFP+ and ChAT-GFP- subsets of GC B cells (CD19+, CD95+, CD38lo) and TFH cells (CD3+, PD1+, Cxcr5+) were sorted from splenic single-cell suspensions. Each replicate consisted of pooled (combined using equal parts), equivalent cell subsets derived from the spleens of two mice to ensure sufficient input cell numbers for RNA isolation. Total RNA was extracted using TRIzol (Invitrogen) according to manufacturer instructions. Library preparation was performed using the NeoPrep Library System (Illumina) following the manufacturer's instructions.