Project description:FAM134B, also known as RETREG1 or JK-1, was first identified in esophageal cancer and later identified as an ER autophagy receptor. The protein contains a Reticulon-homology domain(RHD) and an LC3 interaction region (LIR), where RHD is responsible for sensing and inducing ER membrane curvature, while LIR binds to LC3 on the phagocytic membrane, and the ER membrane and its contents are subsequently encapsulated into autophagosomes and transported to lysosomes for degradation. Now FAM134B, as one of the receptors involved in ER autophagy, has received more and more attention.We knocked down FAM134B in HK-2cells and researched which genes and pathways were been influenced.
Project description:We first performed miRNA analysis on SU-4 cells in suspension, and after 24 and 48 h of HK co-culture. To ensure that no HK cell contamination occurred, SU-4 cells in suspension and after co-culture were purified by CD19-positive magnetic selection, followed by total RNA isolation. Two-condition experiment, S4 vs. S4+HK cells. Biological replicates: 2 suspension controls, 2 co-cultured with HK cells, independently grown and harvested. One replicate per array.
Project description:We first performed miRNA analysis on SU-4 cells in suspension, and after 24 and 48 h of HK co-culture. To ensure that no HK cell contamination occurred, SU-4 cells in suspension and after co-culture were purified by CD19-positive magnetic selection, followed by total RNA isolation.
Project description:Chemotherapeutic use of cisplatin is limited by its severe side effects. In this study, we demonstrated that cisplatin induces cell death in a proximal tubular cell line by suppressing glycolysis- and tricarboxylic acid (TCA)/mitochondria-related genes. HK-2 cells were cultured to confluence in 100mm dishes. Total RNA was extracted (QIAGEN, Valencia, CA, USA), and the concentration in the samples was measured using a Micro UV-Vis fluorescence spectrophotometer (Malcom, Tokyo, JAPAN). Sample of 10ɥg of Total RNA from HK-2 cells were labeled with biotin (3'IVT Labeling Kit, Affymetrix, USA) and hybridized (GeneAtlas Hybridization, Wash, and Stain Kit for 3' IVT Arrays, Affymetrix). The Microarray platform used was Affymetrix HG U219 Array Strip. The arrays were scanned in a GeneAtlas scanner controlled by GeneAtlas Software (Affymetrix, Sta. Clara, USA). Genes were later filtered according to expression fold change (Affymetrix Expression Console software, Affymetrix) Using Affymetrix GeneAtlas System, we determined the gene expression profiles of HK-2 cells treated with cisplatin. Two replicates were made for each timepoints (control, 6h and 24h).
Project description:WWP2 is a E3 ligase which regulates multiple cellular events, such as stress response, differentiation. WWP2 played physiological role in the distal tubules of the kidneys. Whether WWP2 regulated pathophysiological rol in the proximal tubules is unclear. In the present study, we stably overexpressed WWP2 using lenti-viral infection in the human tubular epithelial cells, HK-2, and performed proteinomics analysis to explore WWP2's potential substrates.
Project description:Shotgun time-series proteomic analysis based on stable isotope dimethyl labeling and UHPLC-MS/MSC to relatively quantify the changes in protein expression of human proximal tubular HK-2 cells exposed to a diabetic-like microenvironment
