Fig. 2

Immunoprecipitation of native SjAgo-miRNA-mRNA complex with anti-SjAgo1 antibody. a Western blot assay of immunoprecipitated SjAgo complex. Lane 1 (control): total parasite extract was directly incubated with Protein A agarose without immune serum; Lane 2 (control): pre-immune serum; Lane 3 (control): only immune serum against SjAgo was incubated with Protein A agarose without total parasite extract; Lane4: Protein A-agarose was incubated with immune serum against SjAgo in the presence of total parasite extract. The SjAgo complex and IgG are indicated by asterisks. b RT-PCR products of the small RNAs isolated from the RNA-protein complex after ligation of linkers (17 + 17 nt). cDNA was synthesized according to the manufacturer’s instructions (TaKaRa), then PCR was performed (35 cycles). The putative miRNA and mRNA populations were indicated. c and d Verification of four previously identified S.japonicum miRNAs by PCR analysis. c Lanes 1, 4, 7, 10 are the negative control, lanes 2, 5, 8, 11 are the positive control, and lanes 3, 6, 9, 12 are the experimental groups of cDNAs from Fig. 2b. d Sequencing of the PCR products amplified from the cDNAs from the SjAgo complex