Nucleolar Division in the Promastigote Stage of Leishmania major Parasite: A Nop56 Point of View

Figure 5

Nuclear distribution of Elp3b during the mitotic cell cycle of L. major. Promastigotes were fixed with paraformaldehyde and stained for double indirect immunofluorescence analysis using a mixture of primary antibodies against protein C (for recombinant Elp3b-PTP protein) and β-tubulin (to label subpellicular microtubules and mitotic spindle) followed by anti-rabbit IgG coupled with Alexa Fluor® 594 (red) and anti-mouse IgG conjugated with Alexa Fluor® 488 dye (green) secondary antibodies. DNA was stained with DAPI (blue). L. major promastigotes at early, middle, and late stages of closed mitosis were analyzed. DNA, Elp3b, subpellicular microtubules, and mitotic spindle (indicated with a white arrowhead) are visualized. Colocalization of DNA and proteins is displayed in the Merge column. All images were obtained with a conventional epifluorescence microscope. K: kinetoplast; N: nucleus; No: nucleolus. Size bar denotes 2 μm.