Figure 2.

Consequences of TSA/AzaC treatment on epigenetic status, proliferation and neurosphere-initiating frequency of wt and bcl2-transgenic neurosphere cells. (A) Western blot analysis of untreated and treated bcl2-transgenic neurosphere cells with acetylated histone H4- and MeCP2-specific antibodies. Equal loading was verified by histone H4-specific antibody. (B) [³H]-thymidine incorporation into untreated and TSA/AzaC-treated wt neurosphere cells and TSA/AzaC-treated eGFP/bcl2 neurosphere cells. Means and standard deviations are indicated (n=3). (C) Neurosphere-initiating frequencies of untreated, treated wt and eGFP/bcl2 neurosphere cells. To analyse the effect of TSA/AzaC treatment on neurosphere-initiating frequencies, wt and eGFP/bcl2 neurosphere cells were left untreated or were treated for 48 h with TSA/AzaC. Single-cell suspensions were then prepared and graded numbers of cells were seeded into 96-well plates in NSC medium (n⩾6). Numbers of wells that developed neurospheres 2 weeks postseeding were plotted against the numbers of cells seeded. Data for untreated neurosphere cells are from Figure 1B.