GSM7727624: Nfe2l2fl/fl, scRNAseq; Mus musculus; RNA-Seq
Mice were anesthetized with isoflurane and intranasally administrated with a mixture of 7 ug of PAO and 20 µg of Ova every other day for 9 days (day 0, 2, 4, 6, 8). In house dust mite (HDM)-induced lung inflammation experiments, mice were sensitized on day 0 by 1 µg of Der f 1 via intranasal route. After 1 week, mice were intranasally administered with 10 µg of Der f 1 every day for a total five times (day 7, 8, 9, 10, 11). Sixteen hours after the last administration, all mice were euthanized and bronchoalveolar lavage (BAL) fluid, lung, and mediastinal lymph node were obtained for further analysis. For mixed bone marrow transplantation experiment, donor bone marrow cells from WT control (CD45.1+/+) and Cd4creNfe2l2fl/fl (CD45.2+/+) mice were mixed at 1:1 ratio and adoptively transferred into irradiated recipient Rag1-/- mice. After 4-week reconstitution period, the recipients were intranasally injected with a mixture of PAO/Ova as previously described. To generate single cell Gel Beads-in-Emulsion (GEMs), the 10x Genomics Chromium platform was used to capture and barcode the cells by following the manufacturer's protocol (10x Genomics, Pleasanton, CA, USA). Cell suspensions were loaded on to 10x Genomics Chromium Next GEM Chip K along with the reverse transcription master mix. Along with gel beads coated with oligonucleotides, cells were divided into the GEMs. These oligonucleotides were used for mRNA capture inside the droplet by 13 bp oligo-dT after cell lysis. After reverse transcription, cDNAs with barcodes were amplified and qualified using BioAnalyzer 2100 (Agilent technologies, Santa Clara, CA, USA). The libraries were constructed from these DNA products with the Single Cell Reagent Kit (5' v2 chemistry) for each sample. Adapters were ligated on to the cDNAs and amplified with indexing primer.