title: GSM8213863: PIK3CAWT + alpelisib; Mus musculus; RNA-Seq
library_construction_protocol: 8-week-old PIK3CAWT and PIK3CAPod-HET (PIK3CA gain-of-function mutation specifically in podocytes) mice were treated with vehicle or alpelisib for 4 weeks. 4 mice groups, PIK3CAWT + vehicle, PIK3CAWT + alpelisib, PIK3CAPodo-HET + vehicle, and PIK3CAPodo-HET + alpelisib were processed for scRNAseq profiling to explore the transcriptional programs of PIK3CA overactivation in podocytes, as well as the effect of alpelisib in podocytes. Glomerular enrichment and single-cell suspensions were achieved by using M-450 Dynabeads (Thermofischer Scientific), Multi Tissue Dissociation Kit 1 (Miltenyi Biotec), and size filters (100 um and 70 um). Library preparations was carried out using a Chromium Single Cell 3' Library & Gel Bead Kit v.3 (10x Genomics) according to the manufacturer's protocol. Note that 20,000 cells were loaded in the 10x Chromium Controller. Libraries were sequenced on an Illumina NovaSeq6000 according to the manufacturer's instructions. An equimolar pool of the 4 individual 10X Genomics Single Cell Expression 3' V3 libraries was prepared by the Imagine Genomic Core Facility and sequenced on an S2 FlowCell using the NovaSeq6000, Illumina (sequencing mode Paired-End 100+100 bases + indexes). A total of 2 billion reads were targeted for this pool of 4 libraries (500 million reads per library).