The molecular mechanisms of organ size control and regulation remain one of the major unsolved mysteries of development biology.Almost a decade ago,the discovery of the Hippo signaling pathway in Drosophila shed some light on this puzzling issue.The Hippo signaling pathway is highly conserved in both invertebrates and vertebrates,and plays critical roles in animal development.It controls organ size and growth by inhibiting cell proliferation and by promoting apoptosis.Malfunction of the Hippo signaling pathway leads to cancer development and tumorigenesis.Although the core of the signaling pathway is well understood,the upstream inputs and downstream transcriptional regulation are still obscure to us.In this review,we summarize the current understanding of the mechanism and the function of the Hippo signaling pathway and compare its differences between flies and mammals.We underline the crosstalk between the Hippo signaling pathway and other signaling pathways,and the possible roles of the Hippo pathway in stem cell proliferation and self-renewal.
Recognizing proteins via the production of highly specific monoclonal antibodies(mAbs)is crucial to identifying proteins for proteomic research.However,traditional mAb generation is time-consuming with low efficiency.In this study,we assessed the high throughput method of producing mAbs by immunizing mice with multiple antigens in order to obtain hybridomas against these multiple antigens in one cell fusion.We selected eight proteins that play important roles in human physiological or pathological processes.These proteins were mixed and simultaneously administered to one mouse.We observed the immunizing period for 10 d,and determined the effect of liquid medium and semi-solid medium in hybridoma generation.As a result,all eight immunogens induced antibodies in the immunized mouse in one cell fusion,we obtained hybridomas specific to all eight proteins by enzyme-linked immuno sorbent assay(ELISA)screening,hybridomas against five out of eight showed specific positive in Western-blotting assays.This indicates that we generated mAbs specific to eight proteins in one cell fusion,greatly increasing the efficiency of mAb generation.Furthermore,we observed that hybridomas selected from the liquid medium and semi-solid medium showed different reactivity to antigens.Our study established high-throughput and time-saving methods for production of mAbs.These results provide alternative approaches for increasing the efficacy of mAb generation.
LIU YingWANG YunDanLIU JingZUO WeiHAO LuZHANG LiLiZHEN Bei
