Expression data from mouse splenocytes cocultured with differently treated KLN-205KD cells

Semophorin (SEMA) 6A showed capability to reduce the SEMA3A-derived supression of T cells. However, the effect of SEMA6A on the other immune cells was still unclear. Therefore, we performed gene expression analysis using an Affymetrix mouse genome 430 2.0 array to investigate the potential immune cells regualted by the interaction of SEMA6A and SEMA3A. Thus, we predicted other possible immune cells regulated by SEMA6A using GSEA to analyze the microarray data of co-cultured splenocytes. The reuslts revealed that SEMA6A decreased the SEMA3A-mediated inhibition of B cells and induced activities of dendritic cells in a SEMA3A-indepenet manner. Moreover, M1 and M2 macrophages were not influenced by SEMA6A and SEMA3A. Mouse splenocytes were cocultured with (i) SEMA3A-knockdown KLN-205 cells (KLN-205KD, control line), (ii) KLN-205KD cells overexpressing SEMA6A (SEMA6A), (iii) KLN-205KD cells plus exogenous SEMA3A (SEMA3A), and (iv) KLN-205KD cells overexpressing SEMA6A plus exogenous SEMA3A (SEMA6A+SEMA3A) for 9 days. Interleukin-2 was added at 100 units/mL every three days. Then, The RNA of splenocytes was isoloated to do microarray.