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. 2009 Jul 31;4(7):e6464. doi: 10.1371/journal.pone.0006464

Figure 5. Dimerization of TCTP is necessary for the cytokine-like activity of TCTP.

Figure 5

(A, B) RrTCTP and Del-N35TCTP were irreversibly cross-linked by DMS, BMOE, or BM(PEO)3. Proteins were completely reduced with 20 mM DTT before incubating with BMOE or BM(PEO)3. Crosslinker-treated RrTCTP, Del-N35TCTP, and their 1/2∶1/2 molar mixture were analysed by reducing SDS-PAGE. And, the secretion activities of IL-8 by DTT, BMOE, or BM(PEO)3-treated RrTCTP or Del-N35TCTP were compared using BEAS-2B cells (** p<0.01, ***p<0.001, significantly different from untreated Del-N35TCTP or DTT-treated Del-N35TCTP value, n = 3). (C) Schematic diagram of Fc-fused human recombinant TCTP (Fc-HrTCTP). (D) Fc fusion proteins were analyzed by Western blotting with HRP conjugated anti-rabbit IgG. (E) Fc fusion proteins were analyzed using native gel. The apparent molecular weights were confirmed by the molecular size marker (Sigma). (F) The release activities of IL-8 by indicated protein were compared in BEAS-2B cells (** p<0.01, significantly different from Fc value, n = 3).