RNA-seq for understanding effects of vitamin B12 removal on astrocyte culture

Astrocytes express a vitamin B12 uptake receptor, CD320/TCblR, that is regulated by S1P1 signaling. In search of genes controlled under B12 deficiency in astrocytes, we employed RNA-seq using RNA of astrocytes cultured in normal or B12 deficient media. Overall design: Primary cultures of astrocytes were maintained on either serum-free normal or serum- and B12-free media for 2 weeks and RNA was isolated. RNA-seq was performed starting with 150 ng of RNA from each sample in biological duplicates.