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. Author manuscript; available in PMC: 2011 Feb 7.
Published in final edited form as: Nature. 2010 Aug 5;466(7307):769–773. doi: 10.1038/nature09209

Figure 3. Analyses of L1 silencing in a clonal (pk-5) cell line.

Figure 3

(a) The cartoon indicates the chromosomal location of a silenced L1-retro-EGFP event in a clonal PA-1 cell line. L1-retro-EGFP expression can be reactivated by TSA treatment. (b) Southern blot analysis reveals that pk-5 cells contain a single L1-retro-EGFP event. Genomic DNA was digested with HindIII and the blot was probed with an α-32P radiolabeled EGFP probe. MW=molecular weight standards (kb). (c) Withdrawal of TSA (bottom panels) results in the reestablishment of L1-retro-EGFP silencing. (d) ChIP analysis on naïve and TSA treated pk-5 cells using AcH4 and diMeH3 antibodies. qPCR revealed the enrichment (AcH4) or depletion (diMeH3) of the retrotransposed EGFP sequences in the TSA treated pk-5 cells. The input cycle threshold (Ct) was designated as 1 and used to calculate fold change differences. Samples were run in triplicate from the same experiment. The standard deviation (SD, n=3) is indicated in the graph.