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. 2012 Oct 2;4(10):1112–1125. doi: 10.1002/emmm.201201272

Figure 2. Immunosuppression of STAT1/3/4 phosphorylation by ODNR01 selective binding.

Figure 2

  • A,B. STAT and phospho-STAT Western blot analysis of anti-CD3/28 mAb-stimulated CD4+ T cells treated with indicated cytokines, with and without 5 µM of ODNR01, ODNA151 or ODN1612.
  • C. Confocal microscopy to detect co-localization of FITC-conjugated ODNs (2.5 µM; green) with different Alex Fluor 555-conjugated anti-STAT rabbit polyclonal antibodies (red) in CD4+ T cells after 24 h of incubation.
  • D. Left panels: Immunoblot analysis of STATs from CD4+ T cells pre-incubated with 5 µM biotinylated-ODNs, lysed, and precipitated with avidin agarose beads for STAT detection. Right panels: Immunoblot analysis of STATs from CD4+ T-cell lysates that were pre-incubated with 1 µM biotinylated ODN and 5 µM unlabelled ODN for 1 h and immunoprecipitated with avidin beads for detection of different STAT. The experiments were repeated three times with similar results.