DNA Methylation and Histone Modifications Are the Molecular Lock in Lentivirally Transduced Hematopoietic Progenitor Cells

Figure 8

ChIP assay of CMV promoter region and GFP reporter gene at day 2 and day 7 post-transduction. The chromatin of the non-silenced and silenced HPCs were harvested at day 2 and day 7 post-transduction and immunoprecipitated with anti-histone H3 acetyl lysine 9 (Anti-H3K9) antibody, anti-histone H3 dimethyl lysine 4 (Anti-H3 diMeK4) antibody and anti-histone H3 dimethyl lysine 9 (Anti-H3 diMeK9) antibody. The DNA was subjected to PCR with primers specific to either CMV promoter region or GFP reporter gene and β-actin promoter region (As positive control for ChIP assay). The chromatin harvested without immunoprecipitation (Beads) was set as a negative control for the ChIP assay. The PCR reaction mixture without DNA was set as the negative control for PCR.

(a) CMV promoter region (At day 2 post-transduction)

(b) CMV promoter region (At day 7 post-transduction)

(c) GFP reporter gene (At day 2 post-transduction)

(d) GFP reporter gene (At day 7 post-transduction)