Figure 6. Western blot analysis of subcellular fractionation samples for purity using organelle specific markers

NE-PER reagent was used to isolate nuclear fractions (NF) and post-nuclear fractions (PNF) containing membranes and cytosol from ARPE-19 and COS-7 cells. A differential centrifugation protocol was used to isolate nuclear fractions (NF), membrane fractions (MF), and cytosol fractions (CF) from ARPE-19 cells. Total cell lysates (TCL) were run as positive controls for each condition. The fractions were immunoblotted for PCNA and lamin B as nuclear markers, TGN46 as a membrane marker and α-tubulin as a cytosolic marker.