Single-cell profiling (scRNA-seq with paired V(D)J) of the splenic response to VLP immunization in MZB KO versus wild-type mice
Single-cell profiling (scRNA-seq with paired V(D)J) of the splenic response to VLP immunization in MZB KO versus wild-type mice
Wild-type mice were purchased from Shanghai SLAC Laboratory Animal Co. (Shanghai, China).All mice were housed in a Specific Pathogen Clearance (SPF) grade animal facility with a 12-hour light/dark cycle. Mice had free access to standard autoclaved feed and sterile water. All animal experimental procedures were conducted in accordance with the Guidelines for the Household and Use of Laboratory Animals and were approved by the Animal Ethics Committee.Following the stipulated immunization protocol, mice were humanely euthanized. Spleens were aseptically and surgically resected for subsequent single-cell suspension preparation and analysis. Single-cell suspensions were prepared from mouse spleens. Cell lysis and poly(A) RNA capture were performed simultaneously within the 10x Genomics GEMs, bypassing conventional RNA extraction. According to the manufacturer's instructions, a single-cell RNA-seq library was constructed using the Chromium Next GEM Single-Cell 5' Kit (10x Genomics). Cell lysis and reverse transcription were performed within the GEM to generate barcoded cDNA. The GEM was then lysed, the cDNA was purified, and amplified by PCR to generate the final sequencing library.