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. 2018 Dec 22;47(5):2594–2608. doi: 10.1093/nar/gky1281

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© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Increased piRNA lengths and reduced piRNA levels in adult Tdrkh^cKO testes. (A) Severe reduction of total piRNAs in Tdrkh^cKO testes. Total RNA from adult WT and Tdrkh^cKO testes were end-labeled with [³²P]-ATP and detected by 15% TBE urea gel and autoradiography. (B) The length distribution of small RNAs from adult WT and Tdrkh^cKO testicular small RNA libraries. Data were normalized by miRNA reads (21–23nt). (C) Extension of MILI-piRNAs in Tdrkh^cKO testes. Small RNAs were isolated from immunoprecipitated MILI-RNPs, end-labeled with [³²P]-ATP and detected by 15% TBE urea gel and autoradiography. Western blotting was performed using anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M: molecular weight marker. (D) Diminished MIWI-piRNAs in Tdrkh^cKO testes. Small RNAs were isolated from immunoprecipitated MIWI-RNPs, end-labeled with [³²P]-ATP and detected by 15% TBE urea gel and autoradiography. Western blotting was performed using anti-MIWI antibody to show immunoprecipitation efficiency. M: molecular weight marker. (E) The length distribution of MILI-piRNAs from adult WT and Tdrkh^cKO MILI-piRNA libraries. (F) The length distribution of MIWI-piRNAs from adult WT and Tdrkh^cKO MIWI-piRNA libraries.