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. 2004 Sep;78(18):10104–10110. doi: 10.1128/JVI.78.18.10104-10110.2004

FIG. 3.

FIG. 3.

Detection in vivo of an RNP complex between the CsPP2 and HSVd RNA by immunoprecipitation assays. (A) Phloem exudates (200 μl) were collected from HSVd-infected (lanes 1 and 2) or noninfected (lane 3) cucumber plants and immunoprecipitated with CsPP2 antiserum (lanes 1 and 3). (B) Immunoprecipitates and supernatants were analyzed by SDS-PAGE and stained with Coomassie blue. Note that approximately 50% of the CsPP2 was immunoprecipitated (compare lanes 1 and 3 with lane 2 in panel B). (C) Nucleic acids were recovered from the immunoprecipitate, subjected to RT-PCR with two HSVd-specific primers, and analyzed by PAGE on 5% gels. Samples were blotted and probed with an HSVd-specific riboprobe (D). Lanes C+ and C−, positive (HSVd-RNA) and negative (water) RT-PCR controls. Total nucleic acids were extracted from the immunoprecipitate (IP) obtained in the presence (+) (lane 1) or absence (−) of CsPP2 As (lane 3), protein A (lane 4), or HSVd-RNA (lane 5, uninfected plant). Lane 2 is equivalent to lane 1, except that the phloem exudate was previously UV cross-linked.