Journal: PLoS ONE
Article Title: The Transcription Factor Egr3 Is a Putative Component of the Microtubule Organizing Center in Mouse Oocytes
Figure Lengend Snippet: Egr3 localization is associated with microtubule organization in mouse oocytes. (A) MI oocytes were treated with taxol (microtubule stabilizer, 1 μM) or nocodazole (microtubule depolymerizer, 10 μg/ml) for 1–2 h and were subjected to immunofluorescence staining with anti-Egr3 antibody (Santa Cruz). Red scale bar, 30 μm; white scale bar, 10 μm. Green, Egr3; red, γ-tubulin; blue, DNA. (B) Localization of Egr3 is shown in MI oocyte treated with CytoD (actin depolymerizer, 10 μM) and in Fmn2 -/- oocyte. Fmn2 -/- oocyte was co-stained with anti-Egr3 (Santa Cruz) and anti-γ-tubulin antibodies. Red scale bar, 30 μm. Green, Egr3; red, γ-tubulin; blue, DNA. (C) The localization of Egr3 and microtubule at thawing of vitrified oocytes. Vitrified MII oocytes were stored in LN 2 for 2 weeks. Oocytes were taken out from LN 2 , incubated in decreasing concentrations of sucrose, and then fixed immediately. These oocytes were subjected to immunofluorescence staining with anti-Egr3 (Abcam) and anti-α-tubulin antibodies. Arrows indicate the growing arrays of microtubules at the site of Egr3 accumulation. Green, Egr3; red, microtubule (MT).
Article Snippet: In some experiments, the oocytes were treated with nocodazole (10 μg/ml), taxol (1 μM) or cytochalasin D (cytoD, 10 μM, Sigma-Aldrich) by adding it to the culture media.
Techniques: Immunofluorescence, Staining, Incubation