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Fig. 6. The effect of SRP72 and SRP68 silencing on 7SL RNA localization. (A) Localization of the 7SL RNA by biochemical fractionation. Cytoplasmic and nucleolar fractions were as described in Materials and Methods. RNA from these samples was subjected to northern analysis with random-labeled 7SL and U3 RNA probes from uninduced cells (-Tet) and cells after 3 days of induction (+Tet). (1) SRP72-depleted cells. (2) SRP68-depleted cells. (B) Localization of 7SL RNA during SRP72 and SRP68 silencing by in situ hybridization. Cells uninduced (-Tet) and after 3 days of induction (+Tet) were fixed and hybridized with DIG-labeled PCR probes to U3 and 7SL RNA. (1) Cells carrying the SRP72 RNAi construct hybridized with DIG-labeled PCR probe to 7SL RNA. (2) Cells carrying the SRP72 RNAi construct hybridized with DIG-labeled PCR probe to U3 RNA. (3) Cells carrying the SRP68 RNAi construct hybridized with DIG-labeled PCR probe to 7SL RNA. The nuclei (N) and nucleolus (Nc) are indicated. Bar, 5 µm.
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