GSM5182746: FPWTA1; Mus musculus; RNA-Seq
The tongues dissected from α-gustducin-GFP mice were parted into anterior and posterior. Both anterior and posterior parts were injected with 0.3 mL and 0.1 mL of dispase II  (0.1 U/mL, Sigma-Aldrich) and incubated for 10 min. at 37 ℃, then epithelia surrounding papillae were peeled and incubated in the enzyme mixture containing elastase (0.28 U/mL, Elastin Product) and TrypLE (Sigma-Aldrich) for 15 min. at 37 ℃. The cells were mechanically separated into single cells by glass pipettes (diameter: 100 μm) after filtration using 70 μm cell strainer (Corning). To collect the GFP-positive cells from cell suspensions after cell isolation, FACS was performed using BD FACSAria™ III (BD). Cells were sorted according to cell size (FSC), cell granularity (SSC), 7-AAD (Thermo Fisher Scientific) signals (excitation, 488 nm; emission, 695 nm) for dead cells, FSC pulse-width and SSC pulse-width for singlets, GFP signals (excitation, 488 nm; emission, 530 nm) for GFP-positive cells. For scRNA-seq, cDNA libraries and hashtag libraries were prepared and combined by an BD Rhapsody system and an optimized process similar to our magnetic bead-based bulk RNA-seq method (Shichino et al. JCI insight 2019). Sequencing libraries were generated by using NEBNext UltraII FS library prep kit (New England Biolabs), and QC of cDNA/Hashtag libraries was performed by MultiNA (Shimazu) and qPCR library quantification assay (KAPA). BD Rhapsody WTA
GEO
GSM5182746