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. 2015 Jan 13;25(2):193–207. doi: 10.1038/cr.2015.4

Figure 4.

Figure 4

piRNA-induced mRNA degradation requires the slicer and piRNA-loading activities of MIWI. (A) RT-qPCR analyses of the mRNA levels of Atr, BC026590, Gfpt1, Mdc1, Ppp1cb, Psma8, Tox4, Adam3, Csnk2b, and Fabp9 in enriched Miwi−/− round spermatids relative to those in control Miwi+/− round spermatids. (B, C) Dual-luciferase reporter (B) or qRT-PCR assays (C) of the effect of piR-mmu-7775524 on the expression of BC026590 3′-UTR reporter in GC-2spd(ts) cells. A mutant piR-mmu-77755 (Mut) served as a negative control. (D) Ectopic Flag-HIWI, but not Flag-ADH or Flag-Y345/346A HIWI mutant, restored piR-mmu-77755-induced BC026590 reporter repression in Miwi siR-treated GC-2spd(ts) cells. Data shown represent mean ± SEM of triplicate experiments. **P < 0.01, ***P < 0.001 (Student's t-test). Results shown are representative of three independent experiments.