GSM8499071: FVB.129P2-Pde6b + Tyr c-ch/AntJ control male mice replicate 1; Mus musculus; RNA-Seq
Six (6) pituitaries per sex/diet/transgenic were collected for single-cell RNA-sequencing (scRNA-seq) analyses. Individual pituitary cell suspensions were then pooled within treatment groups (2-3 pituitaries/pool) for counting. Cell concentration within each pool was adjusted to optimally target 10,000 cells according to 10X Genomics guidelines and samples were then submitted to the UAMS Cancer Institute Genomics Core facility. Visual examination under a microscope (EVOS M7000, Thermo Fisher Scientific) was used to detected cell debris or aggregates, which were removed by passing cell suspensions through a Flowmi™ _(SP BEL-ART, catalog # 136800040) cell strainer (40 microns).  Following centrifugation at 250 g for 10 min, cells were re-suspended in 1X PBS (Ca2+/Mg2+-free) containing 0.04% (w/v) BSA. Cell concentration and viability were detected with ReadyProbes™ _Cell Viability Imaging Kit, Blue/Green (Thermo Fisher Scientific, catalog # R37609), and manually counted. Single-cell 3′ library generation was performed using a 10x Genomics Chromium Controller and Chromium next GEM Single Cell 3' Reagent Kits v 3.1 (Dual index). Libraries were sequenced on the Illumina NovaSeq 6000 platform with paired-end mode (read1: 28 cycles, read 2: 90 cycles, i7: 10 cycles, i5: 10 cycles) to generate a minimum of 20,000 read pairs per cell.