Developmental Biology Journal / 2013 / Article / Fig 1

Research Article

Actin Colocalization with Metaphase Chromosomes of the Second Meiosis in Ovulated Mouse Oocytes

Figure 1

Actin distribution in the area of meiosis II metaphase plate. (a–c) Monomeric actin distribution in oocytes stained with Alexa Fluor 488-DNase-I for G-actin and TO-PRO-3 for DNA; (a) actin fluorescence after Alexa Fluor 488-DNase-I labeling; (b) DNA staining by TO-PRO-3; (c) merged image, arrow—metaphase plate, bar −20 μm. (d–f) Double staining for F-actin with TRITC-phalloidin and for DNA with TO-PRO-3; (d) actin stained with TRITC-phalloidin; (e) chromosomes stained with DNA probe TO-PRO-3; (f) merged image, bar −8 μm. (g–i) Double staining for actin by antibody against fragment of N-terminal domain of actin and for DNA by TO-PRO-3; (g) fluorescence of antibody-labeled actin; (h) DNA staining by TO-PRO-3; (i) merged image, bar–24 μm. (j-k) Antibody against fragment of N-terminal domain of actin recognizes microfilaments near metaphase plate; (j) fluorescent staining with antibody, arrows—actin filaments; (k) double staining for actin by antibody and for DNA by TO-PRO-3, arrow—metaphase plate, bar −8 μm. (m-n) After latrunculin B treatment antibody against N-terminus cannot detect actin filaments near chromosomes; (m) fluorescent staining with antibody; (n) double staining for actin by antibody and for DNA by TO-PRO-3, bar −10 μm. (l and o) Disruption of cortical microfilament layer in oocytes after latrunculin B treatment; (l) F-actin stained with TRITC- phalloidin, bar −24 μm; (o) actin fluorescence after labeling by antibody against fragment of N-terminal domain, bar −24 μm.
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