Skip to main content
. 2014 Apr 7;111(16):E1620–E1628. doi: 10.1073/pnas.1321660111

Fig. 5.

Fig. 5.

Association of the 126-kDa protein with the MNase-resistant RNA fragments. TMV-126-F RNA (encodes FLAG-tagged 126-kDa protein) and TMV-126-V RNA (encodes V5-tagged 126-kDa protein) were translated in mdBYL with or without CHX and treated with MNase (1 U/μL). The reaction mixtures without CHX were subjected to immunoprecipitation using anti-FLAG antibody-conjugated agarose [IP (FLAG)]. RNA was purified from the samples before immunoprecipitation (input), from flow-through fractions after immunoprecipitation (FT), and from the eluates from the beads with 3× FLAG peptide (Elu) and analyzed as in Fig. 3A except that a 32P-labeled oligonucleotide probe complementary to nucleotides 11–80 of TMV RNA was used (Upper). The 126-kDa protein was detected by Western blotting using anti–126-kDa protein antibody (Lower). The elution fractions of RNA and protein samples were concentrated 4.3-fold and 75-fold, respectively, compared with the other fractions. Positions of RNA size markers (Upper, in nt) and the FLAG- or V5-tagged 126-kDa protein (Lower) are indicated on the right.