GSM8995841: NTM-infected mouse lung tissue – Atg7 WT (control), scRNAseq; Mus musculus; RNA-Seq
Mouse lung tissues were harvested at 21 days post-infection (dpi) from Mav-infected Atg7 conditional wild-type (cWT) and Atg7 conditional knockout (cKO) mice. The tissues were enzymatically dissociated into single-cell suspensions using Liberase TL (Roche, #05401020001), following the manufacturer's instructions. Cell viability and total cell count were assessed using the LUNA-FX7™ Automated Cell Counter (Logos Biosystems) with Acridine Orange/Propidium Iodide staining. Single-cell suspensions were processed using the Chromium Next GEM Single Cell 3' Kit v3.1 (PN: 1000268) and Chromium Next GEM Chip G Single Cell Kit (PN: 1000120), along with the Dual Index Kit TT Set A (PN: 1000215), according to the manufacturer's protocols (10x Genomics). Single-cell RNA-seq libraries were generated and subsequently sequenced on an Illumina NovaSeq 6000 platform using the NovaSeq 5000/6000 S2 Reagent Kit v1.5 (200 cycles, PN: 20028315). Sequencing was performed in paired-end mode with the following cycle configuration: Read1: 28 cycles, Read2: 90 cycles, i7 Index: 10 cycles, and i5 Index: 10 cycles.