Fig. 5.

Effect of heparin on Δ4 RNA–protein interactions. (A) Polysomes from control (lane C) and ethanol-exposed FCNs (lane E) were incubated with [³²P]-labeled Δ4 sense RNA in the absence of heparin and ethanol-exposed CC polysomes were incubated with [³²P]-labeled Δ4 sense RNA in the presence of heparin (H). At the end of the incubation, samples were processed for RNA gel shift analysis and results were analyzed on PhosphorImager. A shift of Δ4 RNA is seen in the absence of heparin (bracket on the left). Degradation of Δ4 RNA in the presence of heparin into smaller fragments is indicated by open arrows on the right. Free probe (lane P) is indicated by solid arrow on the left. Experiments were repeated three times with similar results. (B) Purified recombinant AnxA2 protein was incubated with [³²P]-labeled Δ4 sense RNA in the absence (lanes 1, 2 and 4) or presence (lane 3) of heparin. Following incubation, samples were processed for RNA gel mobility shift assays. A shift of Δ4 RNA (indicated by open arrow on the right) is observed in the absence (lanes 1, 2 and 4) but not in the presence (lane 3) of heparin. Location of free probe (lane P) is indicated on the left and the addition of heparin to the incubation mix is indicated by (+) below the lanes. Experiments were repeated twice with similar results.