FIGURE 6.

MILI is associated with the cap-binding complex, poly(A)⁺ RNAs, and piRNAs. A, MILI protein co-purifies with components of the cap-binding complex from the testicular extract by the m⁷GTP-Sepharose (lane 3) but not with protein A-Sepharose (lane 2). The interaction is disrupted when m⁷GTP was added to the testicular extract prior to incubation with the m⁷GTP Sepharose as a competitor (lane 4). B, a heterogenous population of poly(A)⁺ RNAs ranging from 0.5 to 4 kb are present in the MILI-immunoprecipitated complex, as detected by oligo(dT)-primed reverse transcription. C, abundant piRNAs are also present in the MILI-immunoprecipitated complex, as detected by direct 5′-end labeling of RNAs extracted from the MILI-immunoprecipitated complex.