Extended Data Figure 4: Drosha-RB-RIIIDmut recognizes stem-loop structures in SINV RNA.
From: RNase III nucleases from diverse kingdoms serve as antiviral effectors
a, Immunoprecipitation of exogenously expressed Flag-tagged proteins. Shown is protein expression in the whole-cell extract and after immunoprecipitation (IP) with a Flag-specific antibody. b, Cells were transfected with Flag-tagged SeV-N or Drosha-RBmut and infected at 36 h.p.t. with SINV at an MOI of 3. At 8 h.p.i., Flag-tagged proteins were immunoprecipitated and bound RNA was isolated to perform qPCR. Graph shows SINV RNA levels relative to input and normalized to tubulin. The average of three independent experiments is shown. Error bars, s.d.; *P < 0.05 using a one-tailed Student’s t-test. c, Prediction of the structure of the 5′ 200 nucleotides of the SINV genome using RNAfold. d, EMSA was performed with the indicated immunoprecipitated proteins and radio-labelled in vitro transcribed RNA comprising the 5′ 200 nucleotides of the SINV genome. Unbound genome is indicated as ‘Free RNA’.
