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Fig. 2. (A) ${gamma}$ -H2AX-containing S-foci partially colocalize with RAD51 foci on SCs. (a) Partial mid-pachytene chromosome spread stained with anti- ${gamma}$ -H2AX (green), anti-RAD51 (red) and anti-SCP3 (blue) antibodies. Examples of non-colocalized ${gamma}$ -H2AX and RAD51 foci are indicated by white arrowheads. Mixed ${gamma}$ -H2AX/RAD51 foci are marked by red arrows. (b-c) Higher magnification of one synapsed chromosome core (b) triple-labeled with anti- ${gamma}$ -H2AX (green), anti-RAD51 (red) and anti-SCP3 (blue) antibodies, (c) double-labeled with anti- ${gamma}$ -H2AX, and anti-RAD51 are shown. ${gamma}$ -H2AX and RAD51 foci are slightly offset to show their colocalization more clearly (d). Red and white arrows indicate sites of colocalized and juxtaposed foci, respectively. (B) ${gamma}$ -H2AX S-foci partially colocalize with MLH1 foci on SCs. (a) Partial pachytene chromosome spread stained with anti- ${gamma}$ -H2AX (green), anti-MLH1 (red) and anti-SCP3 (blue) antibodies. Examples of non-colocalized ${gamma}$ -H2AX and MLH1 foci are indicated by white arrowheads. The only mixed focus on this image is indicated by a red arrow. (b-d) Enlarged images of one SC triple-labeled with (b) anti- ${gamma}$ -H2AX (green), anti-MLH1 (red) and anti-SCP3 (blue) antibodies, and (c) double-labeled with anti- ${gamma}$ -H2AX and anti-MLH1 antibodies. (d) ${gamma}$ -H2AX and MLH1 foci are slightly offset. White arrows indicate juxtaposed foci; red arrows indicate colocalized foci. Bars, 10 µm.

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