NOA36 Protein Contains a Highly Conserved Nucleolar Localization Signal Capable of Directing Functional Proteins to the Nucleolus, in Mammalian Cells

de Melo, Ivan S.; Jimenez-Nuñez, Maria D.; Iglesias, Concepción; Campos-Caro, Antonio; Moreno-Sanchez, David; Ruiz, Felix A.; Bolívar, Jorge

Co-detection of the intracellular localization of polyphosphate and the Flag epitope was performed using a laser TCS-SL confocal imaging system (Leica Microsystems). The Flag epitope was immunostained as described above, using mouse anti-Flag and Cy3-labeled secondary antibodies. Intracellular polyphosphate was detected as described before [39, 43, 44] ; briefly, fixed and washed cells were incubated with 1 mg/ml of DAPI, mounted on slides, and specifically labelled for polyphosphate (excitation: 458 nm, emission: 530-570 nm).