Figure 4

Confocal scanning laser micrographs of (A) human blood cells and (B – D) in vivo matured mouse oocytes labeled with anti-PAD V (N), (E – H) in vivo matured mouse oocytes double labeled with anti-PAD V (N) and LCA, and (I – L) double labeled with ABL₂ and LCA. All anti-PAD V labeling is shown in green, except in A where it is red. DNA stain in A is green. ABL₂ labeling is green and LCA labeling is red in all figures. (A) Cytospin preparations of the granulocyte fraction were stained with anti-PAD V (N), and their nuclei were stained with SYTOX green nucleic acid stain. The merged image shows nuclear localization of PAD (yellow) in a human granulocyte. (B, C) Germinal vesicle intact mouse oocytes and metaphase II oocytes were labeled with anti-PAD V (N), (D) Metaphase II mouse oocyte did not show labeling with goat anti-rabbit IgG conjugated to Alexa 488 alone. (E, F) Polar sections of germinal vesicle intact mouse oocytes double labeled with LCA (red) and anti-PAD V (N) (green). These images were digitally enlarged 2× for better visualization. (G) Merged image of both LCA and anti-PAD V (N) showed co-localization (yellow) of labels. (H) Merged image of equatorial section of metaphase II mouse oocytes double labeled with anti-PAD V (N) and LCA showing co-localization. (I, J) Metaphase II oocytes double labeled with LCA (red) and ABL₂ (green). (K) Merged image of both LCA and ABL₂ showed co-localization (yellow). The inserts of I, J, and K showed the polar view of the oocyte. (L) Control oocytes were not labeled with LCA pre-absorbed with α-D-methyl-mannopyranoside. All samples were imaged at same magnification and the scale bar applies to all figures.