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. 2010 Sep 6;190(5):853–866. doi: 10.1083/jcb.201005117

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© 2010 O’Donohue et al.

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Figure 7. — Nucleocytoplasmic distribution of the 18S rRNA precursors in p-RPS–depleted cells. (A) At 48 h after transfection with siRNAs, cytoplasmic and nuclear RNAs were isolated and analyzed by Northern blotting. A representative experiment shows the patterns obtained after hybridization with the 5′-ITS1 probe for total extracts (To), the cytoplasmic fractions (Cy), and nuclear fractions (Nu). The amount of RNA loaded on the gel was 3 µg/well for total and nuclear extracts and 6 µg/well for cytoplasmic extracts. (B) Percentage of 18S-E pre–rRNA found in the nuclear fraction, as calculated from the balance of the whole fractionation procedure. The levels of 18S-E were measured with a phosphoimager. (C) Amount of 18S-E rRNA in the cytoplasmic fraction relative to control. The amount of 18S-E rRNA in the cytoplasmic fractions was normalized according to the amount of 28S rRNA (measured after hybridization with a specific probe on the phosphoimager), and siRNA-treated cells were compared with control cells. (B and C) The results of two or three independent experiments are shown for each RPS.