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. Author manuscript; available in PMC: 2014 Feb 1.
Published in final edited form as: Mol Microbiol. 2012 Dec 26;87(3):580–593. doi: 10.1111/mmi.12117

Fig. 2. siRNAs co-purify with BB2-LbrAGO1 and are mostly single-stranded.

Fig. 2

A. Pull-down of BB2-LbrAGO1 from LbrAGO1c soluble extract. Aliquots of the input (I), supernatant (S) or pellet (P), each representing 107 promastigote cells, were fractionated on a 10% SDS-PAGE and Western blotted with anti-BB2 or anti-HSP70 monoclonal antibody.

B. The immunoprecipitated BB2-LbrAGO1 material was deproteinized, radiolabeled at the 5’ end and fractionated on a 20% sequencing gel.

C. Purified RNA from the BB2-LbrAGO1 immunoprecipitate was electrophoresed on a 15% native polyacrylamide gel with (lane 1) or without (lane 2) heating at 95°C before loading. The siRNAs were revealed by hybridization to a SLACS oligonucleotide probe. As a control for single- and double-stranded migration, we used a 32P-labeled synthetic 25 nt double-stranded RNA (lanes 3 and 4). ss, single-stranded; ds, double-stranded.