Figure 3: Confocal images of cultured DRG neurons, transiently transfected with PKCI-enhanced green fluorescent protein (EGFP) and PKCβII-EGFP. (a) Cytosolic localization of PKCβI-EGFP. (b) Plasma membrane localization of PKCβI-EGFP. (c) Cytosolic localization of PKCβII-EGFP. (d) Plasma membrane localization of PKCII-EGFP. Method: One-day old DRG cultures were transiently transfected with PKCβI-EGFP (Clontech Laboratories, Inc., USA) or PKCII-EGFP (gift from Dr Yusuf Hannun, Medical University of South Carolina, SC, USA) plasmids using the Effectene Transfection Reagent (QIAGEN) protocol for 12-well culture plates. After 48 h of transfection, the EGFP-positive neurons were visualized by a Confocal Laser Scanning Microscope (TCS SP2 System, Leica) fitted with PL APO 100x/ 1.40-0.7 OIL objective). Sequential scans of EGFP fluorescence were collected and merged and quantified using Leica TCS SP2 Software ver. 3.0.