Notes: The effect of SMYD3 and ERK2 on hepatitis B RNA expression was examine in transfected Huh-7.5 human hepatoma cells. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and converted to cDNA with the GoScript™ Reverse Transcription System followed by dye-based qPCR analysis. The effect of SMYD3 and ERK2 on AP1-dependent and NF-κB-dependent gene expression in the cells was examined with the pGL4.44 [luc2P/AP1 RE/Hygro] and pGL4.32 [luc2P/NF-κB-RE/Hygro] Reporter vectors using the Dual-Luciferase® Reporter Assay System with a pRL-TK Renilla luciferase control. Reporter assays were read with a GloMax® Instrument. (4603)

Notes: To better understand what roles FAM123A may play in signaling, cell behavior and human disease, HT1080 sarcoma cells were plated on MatTek dishes coated with 5mg/ml fibronectin before transfection with Venus (a yellow fluorescent protein)-tagged FAM123A or Venus-WTX, another member of the FAM123 gene family, using FuGENE® HD Transfection Reagent. The cells were imaged the next day for low-resolution analysis. For a higher magnification, dynamic analysis, HT1080 cells were plated onto Delta T dishes coated with 5mg/ml fibronectin and transfected with EGFP-FAM123A and FuGENE® HD Transfection Reagent. The next day, cell images were captured every 10 seconds. Examining the effect of silencing FAM123A, a reporter assay used the pGL4.34[luc2P/SRFRE/Hygro] Vector cotransfected with a CMV-Renilla coreporter and other effector plasmids or siRNA, and luciferase levels assessed using the Dual-Glo® Luciferase Assay System. (4246)