For the purpose of this assignment, I researched an article titled, “The Role of DJ-1 in the Pathogenesis of Endometriosis” by Priyanka Rai and Sisinthy Shivajj. The article discusses many components in which are relevant to the disease, endometriosis. Endometriosis is a disease among women in which causes infertility and severe pelvic pain along with other health concerns. It is seen to occur within 10% of woman whom are of a reproductive or sexually-active age. It is due to the occurrence and growing of the endometrial tissue outside of the uterus, which is abnormal and in result it may cause complications to a woman’s health. Despite research, the underlying cause of endometriosis among woman is uncertain in many aspects. Such as to what women are more prone to the
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Materials and Methods:
This study was carried out at a lab in which involved numerous steps and experimental procedures. The overexpression and reduction of expression of the DJ-1 protein was acted upon normal endometrial cells and endometriotic cells through utilization of siRNA was studied. For the purpose of this experiment, several reagents were used such as: DMEM, FCS, antibiotics, and phenol red free DMEM. There were several cells that were utilized such as: stromal cells, endometriotic epithelial cells, and immortalized human endometrial surface epithelial cells. These cells were then injected with the 200 nM DJ-1 SMARTpool siRNA, which specifically targeted the DJ-1 gene or the non-silencing control. Protein extraction and immunoblotting were then done to prepare the adhesion assay. The cells were lysed and the proteins were eluted through boiling in 3X SDS sample buffer and then the amino concentration was calculated. The proteins were placed in a nitrocellulose membrane, in which they were then ready for immunoblotting. GFP as well as other materials were used as the antibodies for conjugation. The bands were then observed. In order to do the cell
Materials and Methods:
This study was carried out at a lab in which involved numerous steps and experimental procedures. The overexpression and reduction of expression of the DJ-1 protein was acted upon normal endometrial cells and endometriotic cells through utilization of siRNA was studied. For the purpose of this experiment, several reagents were used such as: DMEM, FCS, antibiotics, and phenol red free DMEM. There were several cells that were utilized such as: stromal cells, endometriotic epithelial cells, and immortalized human endometrial surface epithelial cells. These cells were then injected with the 200 nM DJ-1 SMARTpool siRNA, which specifically targeted the DJ-1 gene or the non-silencing control. Protein extraction and immunoblotting were then done to prepare the adhesion assay. The cells were lysed and the proteins were eluted through boiling in 3X SDS sample buffer and then the amino concentration was calculated. The proteins were placed in a nitrocellulose membrane, in which they were then ready for immunoblotting. GFP as well as other materials were used as the antibodies for conjugation. The bands were then observed. In order to do the cell