Effects of supernatants of USP2 knockdown HL-60 cells and mock transfected HL60 cells on differentiated 3T3-L1 cells. Mus musculus

Ubiquitin specifc protease (USP) 2 modulates various cellular responces including carcinogenesis, spermatogenesis, and glucose metabolism. So far, we established USP2 knockdown HL-60 cells to explore roles of USP2 in macrophages. In this experiment, we investigated whether USP2 knockdown HL-60 cells more prominently modulate function of adipocytes compared with control cells. Overall design: USP2 knock down(KD) HL-60 cells and mock vector-transfected mock control cells were differentiated into macrophage-like cells in the presence of 30nM of phorbol 12-myristate 13-acetate for 5 days. Subseqently, supernatants of the USP2 KD cells and control cells were added to differentiated 3T3-L1 cells for 3 and 6 hrs. Total RNA was isolated from the supernatant-treated 3T3-L1 cells, and subjected to microarray analysis. Duplicate samples were prepared for each cell and time point.