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Fig. 2. (a-d) Immunostaining of Rad51 (green). No Rad51 (a) or very little (b) was detected in the early stage II MIC. (c) A large amount of Rad51 was detected in the somewhat later stage II MIC. (d) A stage IV MIC with granular staining of Rad51 (projection of a deconvolved Z stack of images). (e-l) Immunostaining of Rad51 (green) and phosphorylated histone H3 (phosH3) (red). phosH3 acts as a marker for chromatin condensation. (e) MACs and MICs in a conjugated pair of cells with slightly asynchronous meioses. In the smaller (earlier) MIC (left), the red phosH3 staining predominates because there is only a small amount of Rad51 present. In the later MIC (right) the strong Rad51 and the phosH3 staining mix to produce a yellow color. (f) In this stage III MIC, both phosH3 and Rad51 produce granular immunostaining patterns but the spots do not overlap. (g-l) Stages IV (g), V (h), metaphase I (i), anaphase I (j), anaphase II (k) and the tetrad stage (l). Some Rad51 remains on chromatin (overlapping with phosH3) until anaphase I, but much of it is shed from the chromosomes by metaphase I (note the green staining of the spindle in i), and it has virtually disappeared by anaphase II. Note that the MACs contain Rad51-positive spots (see text). The chromatin is stained with DAPI (blue). Bars, 5 µm (in f for a-f) and 10 µm (in l for g-l).
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