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. 2000 Jan 15;28(2):582–592. doi: 10.1093/nar/28.2.582

Figure 4.

Figure 4

Quantitative analysis of the characteristics of nucleocytoplasmic shuttling of oligonucleotides. (A and B) Analysis of P=S ODN 12182–Texas Red shuttling. (C) Analysis of phosphodiester 2′-O-propyl-oligoribonucleotide (PO-2′P-ON) 12183–Texas Red migration. Oligonucleotides were injected at a concentration of 30 µM into one nucleus of binucleate Ref52 cells treated as indicated below. Co-injection of 70 kDa dextran–Cascade Blue marked the injected nucleus. Two hours after injection cells were fixed and the fluorescence intensities per unit area in each nucleus were measured. Then the ratio between the values of the non-injected and the injected nuclei was calculated (see Materials and Methods). In each case at least 10 cells of at least two independent experiments were evaluated. The means ± SD are given. The extent of shuttling was compared for untreated cells (w/o), chilled cells (4°C), energy-depleted cells (–ATP), cells pre-injected with wheatgerm agglutinin (WGA), cells pre-injected with WGA and then injected with 30 µM 12182–Texas Red together with 2 mM unlabeled P=S ODN 8424 (WGA+2mM 8424) and cells that received 12182–Texas Red together with 1.7 mg/ml of a Ran mutant protein impairing nuclear protein export (RanT24N).