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. 2010 May 12;285(29):21988–21993. doi: 10.1074/jbc.M109.041046

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — The transfer of HCs may occur directly between HA sites. HC·HA molecules were generated by incubation of IαI, TSG-6, and HA. The bikunin-containing proteins were subsequently removed by anhydrotrypsin-Sepharose affinity absorption. The resulting sample was analyzed directly (lane 1), treated with NaOH (lane 2), incubated with oligo-HA 10EF and TSG-6 (lane 3), or incubated with oligo-HA 10EF and TSG-6 and subsequently treated with NaOH (lane 4). For comparison, IαI treated with NaOH is also shown (lane 5). The proteins were resolved by reduced SDS-PAGE and immunoblotting using HC1/HC2 antisera. The observed weak bands around 120 kDa are likely to correspond to HC·TSG-6 complexes (lane 3). The band shift of the HCs after incubation with oligo-HA 10EF and TSG-6 (lane 3), when compared with the NaOH-treated samples (lanes 2, 4, and 5), demonstrate that HCs are transferred from HA to HA without the presence of bikunin proteins.