Figure 1. Cytoplast generation and... | Rockefeller University Press

Figure 1.

$Figure 1. Cytoplast generation and characterization. (A) Illustration of enucleation procedure. (B) Fluorescence images of plated fractions 2 h after enucleation. (C) Cytometric profiles of stained populations with a fluorescent nuclear dye (Vybrant DyeCycle Green). Q2 is region containing positive nuclear staining. Q3 is negative for nuclear staining. (D) Western blots of intact cell, cytoplast, and nucleoplast fractions. (E–G) Immunofluorescent staining for nuclear proteins (E), cytoskeletal elements (F), and organelles (G). Arrowheads in G mark centrosomes. All nuclei are Hoechst stains and shown in red. Cell outlines are white. (H) Cell population as percentage of starting population over time shown as mean ± SEM (n = 4 experiments). Student’s t test performed between successive time points for either intact cells or cytoplasts. ***, P < 0.001; **, P < 0.01; *, P < 0.05. Bars, 50 µm.$

Cytoplast generation and characterization. (A) Illustration of enucleation procedure. (B) Fluorescence images of plated fractions 2 h after enucleation. (C) Cytometric profiles of stained populations with a fluorescent nuclear dye (Vybrant DyeCycle Green). Q2 is region containing positive nuclear staining. Q3 is negative for nuclear staining. (D) Western blots of intact cell, cytoplast, and nucleoplast fractions. (E–G) Immunofluorescent staining for nuclear proteins (E), cytoskeletal elements (F), and organelles (G). Arrowheads in G mark centrosomes. All nuclei are Hoechst stains and shown in red. Cell outlines are white. (H) Cell population as percentage of starting population over time shown as mean ± SEM (n = 4 experiments). Student’s t test performed between successive time points for either intact cells or cytoplasts. ***, P < 0.001; **, P < 0.01; *, P < 0.05. Bars, 50 µm.