RNA-seq analysis of gene expression regulated by LSD1 in CD8+ T cells

In the RNA-seq analyses of in vitro activated and expanded CD8+ T cells, we found that GSK treatment significantly altered the transcriptome, with 241 genes upregulated and 426 genes downregulated respectively . Gene Ontology (GO) enrichment analysis revealed that the upregulated genes were highly enriched in biological processes related to cell killing and cytokine production. Mouse CD8+ T cells were isolated from the spleens and lymph nodes of the mice using a biotin selection kitwith an EasySep™ Mouse CD8+ T Cell Isolation Kit according to the manufacturer’s manual. The CD8+ T cells were stimulated with 2 μg/ml plate-bound anti-CD3 (2μg/ml) and 2 μg/ml soluble anti-CD28 (2μg/ml) antibodies with the addition of 20 ng/mL hIL-2 at a density of 1 million/ml for 48 h. Activated CD8+ T cells were expanded with hIL2 in fresh medium for the indicated time before analysis. 0.5 μM GSK2879552 or vehicle control was added. Total RNA was extracted and used to poly(A)+ RNA enrichment by magnetic mRNA isolation kit for constructing library. The prepared libraries were sequenced at 150 bp from both ends.