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The organization of the 5′ portion of Dicer locus is depicted. The targeting vector used for introduction of FlagHA2 into the Dicer locus and the schematic map of the targeted Dicer gene before and after Cre mediated-recombination are shown. Triangles represent loxP sites as indicated. Rectangles indicate the position of Neomycin (Neo) and Diptheria toxin A (DTA) selection marker genes. The SacI restriction sites are indicated as well as the respective Southern fragments detected by the 3′probe. A schematic diagram of the resulting FlagHA2-Dicer protein is shown. (B) Southern blot of tail derived SacI-digested DNA from wild-type and Dcr+/FH-Neo mice is shown with the 3′ probe indicated in A. (C) Western blot using anti-HA and anti-SMC1 antibodies on extracts from adult wild type and Dcr+/FH testis is shown. (D) Immunofluorescence using anti-HA and anti-γH2AX antibodies on Dcr+/FH testis germ cells from adult testis sections is shown. Scale bar = 10 µm. (E) Hematoxylin and eosin stained testis section from adult DcrCtl and DcrC-KO mice with representative tubules shown. Scale bars = 50 µm and 20 µm in the upper and lower panel, respectively. (F) Increased apoptosis in DcrC-KO testis. A TUNEL assay counterstained with DAPI is shown on testis sections from adult DcrCtl and DcrC-KO mice. The apoptotic cells stain in green. Scale bars = 50 µm and 10 µm in the upper and lower panel, respectively. Abbreviations: P, pachytene and RS, round spermatid. 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