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. Author manuscript; available in PMC: 2012 Oct 22.
Published in final edited form as: Methods. 2010 Jul 6;53(1):91–96. doi: 10.1016/j.ymeth.2010.07.001

Figure 4. Detection of methylated Tat in cells.

Figure 4

(A) Immunoprecipitation/western blot analysis of Tat/FLAG coexpressed with KMT1E/FLAG or KMT7/FLAG in 293T cells. Cells were transfected with 1 μg of EF-1α-Tat/FLAG expression vector and 6 μg of pCDNA3.1-KMT1E/FLAG, pCDNA3.1-KMT7/FLAG or empty vector. Lysates were immunoprecipitated with α-FLAG M2-agarose, eluted with FLAG peptide and analyzed by western blotting with α-FLAG, α-me1ARM, α-me2ARM or α-me3ARM antibodies.

(B) Western blot analysis of whole cell lysates after transfection with constructs expressing KMT1E/FLAG or KMT7/FLAG in 293T cells. Cells were transfected with 6 μg of pCDNA3.1-KMT1E/FLAG, pCDNA3.1-KMT7/FLAG or empty vector. Whole lysates were analyzed by western blotting with α-FLAG, α-H3K9me2, α-H3K9me3, α-meH3K4me1, α-H3K4me2, α-H3 or α-tubulin antibodies.