Figure 2. Detection and sub-cellular distribution of PYK2 and FAK kinases.

Western blot analysis of MII oocytes (left) was performed on groups of 60 oocytes/ lane probed with antibodies to FAK protein (FAK protein), antibodies to phosphorylated FAK (FAK PY⁸⁶¹), anti PYK2 protein (PYK2 protein), or anti phosphorylated PYK2 (PYK2 PY⁵⁷⁹) as described in’ Materials and Methods’. The apparent molecular weight of each major band was calculated by comparison with molecular weight standards and is presented in the margins (arrows).

Sub-cellular localization of FAK and PYK2 protein (right) was performed on oocytes collected prior to fertilization in vitro (A, C) or during anaphase/ telophase II (B, D) then labeled with anti-PYK2 protein and anti-FAK protein antibodies not targeted to phosphorylation sites in order to establish the sub-cellular distribution of the entire pool of these kinases in the oocyte. Bound antibodies were detected with alexa 488-anti-rabbit IgG (green). Chromatin was stained with ethidium homodimer (red) or Hoechst 33258 (blue). Magnification is indicated by the bar which represents 10 μm.