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. 2010 Nov 1;21(21):3722–3734. doi: 10.1091/mbc.E10-08-0693

Figure 7.

Figure 7.

Rlg1 proteins bind both HAC1 mRNAs and intron. (A) HA-ScRlg1p and HA-AtRlg1p[M74] were immunoisolated in the presence (Mg2+) or absence (EDTA) of Mg2+ from Tm-treated cell extracts with anti-HA-agarose. RNAs in each fraction were subjected to Northern blotting with the anti-HAC1u probe. T, one-tenth of total; U, one-third of unbound fraction; B, all of bound fraction. (B) HA-AtRLG1[M74] extracts were subjected to discontinuous sucrose density gradients (16/50, 18/50, or 20%/50%) to separate the nonribosomal (N) and the ribosomal (R) fraction described schematically in the upper region. Left, a urea-PAGE image of low-molecular-weight RNAs in the N, R, and T (total) fractions of sucrose density gradients. Right, the extract from Tm-treated AtRLG1 cells was separated to nonribosomal and ribosomal fractions with the 16/50% sucrose density gradient. RNA samples from these fractions were subjected to IP with anti-HA-agarose. Coimmunoprecipitated RNAs were analyzed as in A. Recoveries of HAC1 RNA species in the bound fractions were shown in the bottom of the Northern images.