Figure 5.

B7-H4 promotes the growth of HEK293 cells in vitro and accelerates G1/S phase progression in HEK293 cells. (a) Growth curves of mock/293, B7-H4 WT/293 and B7-H4 H250Q MT/293 cells measured by CCK8. Equal numbers of each cell type were incubated in medium containing 5% fetal calf serum. At 24, 48 and 72 h, CCK8 were added and OD 450 was detected. Data represents the fold difference in OD 450 compared with that at 0 h. Results are mean±s.e.m. from two separate experiments. (b) The fluorescence-activated cell sorting (FACS) analysis indicated that B7-H4 WT promoted G1/S phase transition, while B7-H4 H250Q MT did not. (c) B7-H4 WT could upregulated the expression of Cyclin D1 and Cyclin E in 293 transfectants. (β-Actin was used for testing the house keeping protein for confirming equivalent loading). (d) Mock/293, B7-H4 WT/293 and B7-H4 H250Q MT/293 cells were mixed with Matrigel and subcutaneously injected into contralateral side of the mice as described in method. Aspect of the tumors in vivo and before dissection in the representative mice of three groups were photographed. (e) 24 Mice were divided into three group (8 mice per group). Cells were injected into right flank. After 8 weeks mice were killed and tumors were dissociated. Tumor volumes on 8th week were evaluated as described in method. (f) Tumor weight on 8th week was measured.Data was analyzed by SPSS program.