Cellular localization of the GSE24.2 peptide containing nuclear localization signals at the N-terminal region.

pcDNA3 vectors were generated coding for the GSE24.2 peptide fused to the NLS1 nuclear localization signal (KRKR) or the NLS2 signal (KKEKKKSKK) fused to the N-terminal region, named 24.2-NLS1-5 and 24.2-NLS2.5 respectively. The nuclear localization signals were placed between the N-terminal 6xmyc epitope and the GSE24.2 peptide in the proteins expressed from these vectors. HeLa cells were transformed with the pcDNA3 vectors expressing GSE24.2, 24.2-NLS1.5 and 24.2-NLS2.5, fixed, permeabilized and incubated with a c-myc antibody. Immunofluorescence staining was observed after incubation of the preparations with a secondary antibody conjugated with Alexa fluor 488 using a Nikon 90i microscope. (PDF)