Figure 3

A two steps PCR recombination assay is used to identify Vk to Jk1 or Vk to Jk2 rearrangements from pre-B cells upon RAG induction with Imatinib or GSK. (A) A Sybr Green stained 1.5% Agarose TBE gel in which the recombination PCR reactions templated with initial 1:5 dilutions of genomic DNA extracted from each distinct cell treatment lot (2 × 10⁶ cells) are electrophoretically resolved. The cells were either unexposed (gel reaction lanes 4 and 5) or subjected for 48 h to 1 GHz EMF irradiation (lanes 1 to 3 and 6 to 8) with the generator setting at 13 dBm. The color code designating the positions of exposed EMF (exp.Well) wells in the dish is the same with the one used in supplementary Fig. 1Sb. Last lane (9) of the gel, -DNA control reaction. The bottom black box (cropped from a distinct gel) displays Hisone H1 PCR reactions templated with the same amount of genomic DNA as the recombination reactions above(control genomic DNA). (B) Identified Vk to Jk2 recombined products were quantified from scanned gels corresponding to PCR reactions from cells +/− Irradiation and the calculated ratios of band intensities expressed + EMF/−EMF(irradiated/nonexposed) for each well (color code consistent with that shown in Fig. 1S). The histograms represent the average values of three independent quantified experiments. EMF-Electromagnetic Field, Recombination pharmacological stimuli (Imatinib, IMA) versus (GSK-690693, GSK). H1, histone H1 control reaction PCR reactions. Darker font histograms correspond to lower 7 dBm (l) and brighter to higher 13 dBm(h) generator power settings.