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. 2012 Jun 22;7(6):e39729. doi: 10.1371/journal.pone.0039729

Figure 7. SAM68 is involved in the miRNA pathway in male germ cells.

Figure 7

(A) Total extract from secondary spermatocytes were immunoprecipitated with an anti-SAM68 antibody and analysed in Western blot with anti-DICER and anti-Sam68 antibodies. (B) Cellular extracts from secondary spermatocytes (II sp.cytes) were immunoprecipitated with an anti-DROSHA antibody and detected with anti-DROSHA, anti-SAM68 and anti-hnRNP A1 antibodies. (C) Western blot analysis of co-immunoprecipitation performed in the presence of RNase in the cell extract. (D) Schematic representation of the microRNAs differentially expressed in the indicated Sam68 knockout germ cells. In green are indicated the down-regulated microRNAs, in red the up-regulated ones. The immunofluorescence images show the localization of SAM68 in the corresponding wild type germ cells. (E) Real Time PCR analysis of miR-720, miR-142-3p and miR-29b in RNA purified from round spermatids of Sam68 wild-type (black) or knockout (grey) mice. The values were normalized against U6 snRNA and expressed as fold increase with respect to the value of wild type cells.