title: Sample 9
organism: Mus musculus
description: Protocols: Inguinal lymph nodes were harvested, collected into RPMI 1640 medium containing 2% FCS, 20mM Hepes (all from Lonza), 0.375 mg/ml Collagenase P (Sigma Aldrich) and 100 μg/ml DNase1 (Applichem) and shred into small pieces using 22 guage needles. Samples were incubated at 37C for 60 min, with resuspension and collection of the supernatant every 15 min into PBS containing 0.5% FCS and 10mM EDTA. Stromal cells were enriched by depleting CD45+ hematopoietic cells and Ter119+ erythrocytes using MACS microbeads (Miltenyi). VSV immunization protocol: Mice were immunized sub-cutaneously (s.c) with 2x10E7 PFU UV-inactivated vesicular stomatitis virus (VSV) on day 0 and boosted with a subsequent s.c. immunization with 5x10E7 PFU UV-inactivated VSV on day 4. UV-inactivation was performed by exposing virus to UV light for 20 minutes. Reticular cells / germinal center B cells from draining inguinal lymph nodes were examined at day 12 following the first VSV immunization. EYFP+ TdTomato+ cells from lymph nodes of naïve Cxcl13-Cre/TdTomato EYFP mice and immunized Cxcl13-Cre/TdTomato EYFP or Cxcl13-Cre/TdTomato Cxcl12fl/fl EYFP mice were sorted using the BioRad S3 Cell Sorter or the BD FACS Melody cell sorter. Single cell suspensions were run on the 10x Chromium (10x Genomics) system. The cDNA library generation was performed following the established commercial protocol for Chromium Single Cell 3' Reagent Kit (v2 Chemistry)
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-01-13T04:04:19Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-10-17T12:32:41Z
attributes.tag: External Id
attributes.value: SAMEA6096709
attributes.tag: INSDC center name
attributes.value: Institute of Immunobiology, Kantonsspital St. Gallen, St. Gallen, Switzerland
attributes.tag: INSDC first public
attributes.value: 2020-01-13T04:04:19Z
attributes.tag: INSDC last update
attributes.value: 2019-10-17T12:32:41Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8445:Sample 9
attributes.tag: age
attributes.value: 6 to 8
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: Tg(Cxcl13-Cre/TdTomato)+ x R26-EYFP++
attributes.tag: individual
attributes.value: 11 mice pooled
attributes.tag: infect
attributes.value: UV-inactivated vesicular stomatitis virus
attributes.tag: inferred cell type
attributes.value: dendritic cell
attributes.tag: organism part
attributes.value: inguinal lymph node
attributes.tag: sample name
attributes.value: E-MTAB-8445:Sample 9
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: mixed
attributes.tag: strain
attributes.value: C57BL/6N