GSM8942283: HK-390 cells MXC-017 treated; Homo sapiens; RNA-Seq
At day 5, the gliomaspheres in the suspension culture were collected and dissociated with TrypLE (no phenol red), and the adherent differentiated cells were washed with HBSS and de-attached with Trypsin/EDTA. The cells were then pooled and filtered through a 40-µm strainer and counted with a hemacytometer. The samples were pelleted by centrifugation at 300 g for 5 minutes at 4°C, resuspended in the provided Cell Resuspension Buffer (Fluent Bioscience, Watertown, MA), and 50,000 cells were targeted for capture and processed for single-cell transcriptomics using the PIPseqTM T2 3' Single Cell RNA Kit v4.0 Plus (Fluent Biosciences). Sequencing libraries were prepared according to PIPseq T2 instructions, with 12 cycles for cDNA amplification and 8 cycles for library amplification, depending on the DNA amount. The libraries were quantified using a Qubit 4 Fluorometer and a TapeStation 4200 and sequenced as per manufacturer recommendation on a NovaSeq X Plus – 25B 2 x150bp, with 3.25 billion reads in total for 8 samples.