Fig. S5.

Absence of pachytene piRNAs in the brain. (A) Absence of Miwi in the adult brain. Cytosolic fractions from 8-wk-old hippocampi or PFC were immunoprecipitated and Western blotted using anti-Miwi antibodies. Brain tissues were used at 100 and 1,000 times more than adult testes for immunoprecipitation. Mili^−/− testes, which were also devoid of Miwi expression (22), were used as a negative control for the anti-Miwi antibody. (B) Absence of pachytene piRNAs (Miwi-associated) in the neurons. Reads with 5′U from neuronal small RNA libraries on removal of miRNAs, structural RNAs, and repeat- and gene-derived sequences, were mapped to pachytene clusters as described (19). A reference to the pachytene clusters in the adult testes was made for comparison, by mapping adult testes small RNAs from testes small RNA library as described above.