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. 2018 Dec 28;6(3):455–468. doi: 10.1093/nsr/nwy163

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© The Author(s) 2018. Published by Oxford University Press on behalf of China Science Publishing & Media Ltd.

This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

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Figure 5. — DAZL recruits PABP to regulate translation of its target. (A) Sucrose density gradient fractionation of 10dpp mouse testes with and without DAZL. (B) Immunoblot analysis of different fractions. (C) Target mRNAs from polysome fractions decreased dramatically with Dazl KO. (D) Silver staining of SDS-PAGE gel of DAZL and IgG immunoprecipitation (IP) in 8dpp mouse testes. Proteins were identified by mass spectrometry from excised bands (the whole lane except heavy and light chain). (E) Silver staining of SDS-PAGE gel of DAZL and IgG IP in 25dpp mouse testes. Proteins were identified by mass spectrometry from excised bands (the whole lane except heavy and light chain). (F) Gene ontology analysis results of proteins identified by mass spectrometry from (D). GO:0000028: ribosomal small subunit assembly; GO:0001649: osteoblast differentiation; GO:0000387: spliceosomal snRNP assembly; GO:0021762: substantial nigra development. (G) Gene ontology analysis results of proteins identified by mass spectrometry from (E). GO:1904874: positive regulation of telomerase RNA localization to Cajal body; GO:0032212: positive regulation of telomere maintenance via telomerase; GO:1904871: positive regulation of protein localization to Cajal body; GO:1904851: positive regulation of establishment of protein localization to telomere. (H) Co-IP of DAZL and PABP in 8dpp mouse testes. (I) Co-IP of DAZL and PABP in 25dpp mouse testes. (J) Biotin-oligo was designed to reverse complement the 3′UTR of Sycp1 mRNA. (K) 3′UTR of Sycp1 mRNA was enriched in Sycp1 pull-downed samples. (L) DAZL and PABP proteins were enriched in Sycp1 pull-downed samples. (M) Overexpression (OE) of DAZL in SSCs enhanced the expression of PLZF and LIN28A. Relative expression of PLZF and LIN28A in different samples was quantified using ImageJ.