Biotin end-labeled oligonucleotides were conjugated to Dynabeads M-280 Streptavidin (Thermo Fisher Scientific, 11205D) for 30 min at room temperature according to the manufacturer’s protocol. Conjugated beads were incubated for 2 hours at 4°C with cleared lysates from confluent cells transfected with the appropriate overexpression plasmids. The beads were then washed four times with wash buffer, and the bound proteins were eluted by 95% formamide, 10 mM EDTA, and free biotin at 65°C for 10 min and then analyzed by Western blotting. The oligonucleotides used are as follows: hIL6 3′UTR, 5′-biotin-UAUUUUAAUUAUUUUUAAUUUA-3′; 2×GAS, 5′-biotin-CGTTTCCCCGAAATTGACGGATTTCCCCGAAAC-3′ (forward) and 5′-GTTTCGGGGAAATCCGTCAATTTCGGGGAAACG-3′ (reverse); nonGAS, 5′-biotin-CGTTTACCCCAAATTGACGGATTTACCCCAAC-3′ (forward) and 5′-GTTGGGGTAAATCCGTCAATTTGGGGTAAACG-3′ (reverse); newMOTIF, 5′-biotin-CGTTTTTGAGGCAATTGACGGATTTTTGAGGCAAC-3′ (forward) and 5′-GTTGCCTCAAAAATCCGTCAATTGCCTCAAAAACG-3′ (reverse). All oligos were purchased from Thermo Fisher Scientific.

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