GSM8988018: wt BMDM UT; wt BMDM 2hLPS; Smarca5KO BMDM UT; Smarca5KO BMDM 2hLPS; GEX; Mus musculus; RNA-Seq
Single-cell RNA sequencing was performed using the 10x Genomics Chromium Next GEM Single Cell 3' Kit v3.1, following the protocol (CG00391 Rev B) provided by the manufacturer. Cultured cells were first diluted in PBS, washed, and labeled using Cell Multiplexing Oligos (10x Genomics, 3' CellPlex Kit Set A 1000261), in accordance with the manufacturer's guidelines. After labeling, cells were washed again, counted to ensure a viability above 80% and minimal aggregation, and then prepared for loading. Labeled cells were pooled, recounted, and loaded onto a Chromium Next GEM Chip G using the Chromium Controller (10x Genomics). Reverse transcription, cDNA amplification, and library preparation, were carried out as per the manufacturer's protocol (CG000388). Library quality was assessed using the Agilent TapeStation HSD5000 ScreenTape system, and quantification was performed with both the Quantifluor Fluorometer (Promega). Sequencing was conducted on an Illumina NovaSeq6000, with paired-end reads (28 bp Read 1 and 90 bp Read 2).