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. Author manuscript; available in PMC: 2014 May 6.
Published in final edited form as: Leukemia. 2013 Jun 14;27(9):1832–1840. doi: 10.1038/leu.2013.180

Figure 2. Mutational analysis and identification of TLR2-F217S in MDS.

Figure 2

(a) Representative Sanger sequencing traces showing nucleotides around coding region of TLR2-F217 in whole bone marrow (WBM) DNA (Top) and CD3+ genomic DNA (Bottom) of one MDS sample. (b) Schema of TLR2 gene and sequence alignments between human and other species. (c) Western blots of GFP fusion protein expression of TLR2 WT and F217S in 293T cells. (d) Luciferase analysis of activation of NF-kB by WT and F217S TLR2 in transfected 293T cells. Cells were exposed to either no agonist or to PAM2CSK4, PAM3CSK4 or MALP2 known TLR2 agonists. Pooled data from three separate experiments. (e) Western blot characterization of IRAK1 modification in 293T cells transfected with either WT or F217S TLR2 and treated with no agonist, MALP2 or PAM2CSK4. Top, Western blot analysis of IRAK1; Middle, IRAK1 immunoprecipitation followed by phospho-IRAK1 Western blot; Bottom, IRAK1 immunoprecipitation followed by poly-ubiquitin (K63).