L lp - hsdl ma of i . 5kb was detected in the rna extracted from cult ' ured chorionic trophoblast using northem blot hybridhation method . the expression of ll6 - hsdl wa was induced by treating the chorionic cells with dexamethasone ( l0 - ' , l0 ' m ) for 24h , and the induction was blocked by co - treatinent of the cells with ru486 . these results suggest a gr - mediated effect of glucocorticoid on the rnrna expression and reductase activity of l l 9 - hsd1 人h合成的肌?一加x门’ m )对培养的绒毛膜滋养层细胞的11p hsdi还原酶活性具有诱导作用,此诱导作用可以被gr阻断剂ru486门0川)所阻断; northern印迹杂交方法进一步证实, dex亦可诱导绒毛膜滋养层细胞11p叫inrna的表达,此诱导作用也可以被ru486门0叫川)所阻断
In this study , we showed that the pdk inhibitor ly294002 regulated the cellular localization of materials 1 . kunming mice were supplied from the department of laboratory animals , china medical university 2 . pregnant mare serum gonadotropin ( pmsg ) and human chorionic gona - dotropin ( hcg ) were obtained from tianjin huafu biological products research institute and shanghai products research insititute , respectively 为了研究在小鼠受精卵中pkc 、 pkb是否也通过p21蛋白参与调控g2 m转换,本实验应用了pma和ly294002作为pkc 、 pkb的抑制剂,研究它们对小鼠受精卵中p21蛋白表达和定位,进一步探讨可能的调控机制。
The co - expression of l l6 - hsd1 and gr in the saxne chorionic trophoblast suggests possible intrcrine achons ofglucoconicoid generated by l l6 - hsdl within the cells . 2 . ewe radiomctric conversion assay showed that trctrient of the cells with the synthetic glucocorticoid - - dexarnethasone ( l0 - ' m ) for 24h increased the conversion of conisone to cortisol , and thes increase was blocked by the gr antagonist ru486 双标免疫组织化学染色结果显示11p hsdi和gr共存于同一个滋养层细胞,提示在体内无活性的gc代谢产物?一17羟d脱氢皮质酮经11p hsdi还原活化后可以直接与同一个细胞内的gr结合,即以内在分泌( intracrine )形式发挥作用
Anti - p21 mouse monoclonal antibody from beijing zhongshan biotechnology anti - mouse or anti - rabbit igg secondary antibody from santa cruz biotechnology ly294002 from sigma biotechnology tritonx - 100 from boehringer mannhein gmbh fluorescein isothiocyanate ( fitc ) conjugated anti - mouse igg antibody was purchased from beijing zhongshan biotechnology hepes from e . metck darmstadt methods superovulation and collection of eggs for superovulation , female kunming mice 4 - 5 week old were injected with pregnant mare serum gonadotropin ( pmsg ) , and after 46 - 48 hours with human chorionic ginadotropin . ( hcg ) . one - cell fertilized eggs were collected on the next day from oviduct of females 取4一5周龄成熟雌性昆明系小白鼠,腹腔注射pmsg (孕马血清促性腺激素) 10iu , 46一48小时后腹腔注射hcg (人绒毛膜促性腺激素) 1oiu ,将注射hcg后的雌鼠与8周以上的成熟雄鼠合笼交配,次日检察阴栓,将查到阴栓的雌鼠处死,取输卵管于mz培养液中,解剖镜下撕开壶腹,释放细胞团,然后用300林岁nil透明质酸酶消化去除颗粒细胞,口控吸管将卵细胞在m :中反复清洗,然后置于孵箱中,根据时间点收集g2期细胞。