Figure 3

The expression of meiosis-specific genes DMC1, REC8, STAG3, SYCP1 and SYCP3 in irradiated lymphoma cells in the time-course post-irradiation: (A) Transcription of REC8, STAG3 and DMC1, shown by RT-PCR of mRNA isolated from WI-L2-NS, TK6 or Namalwa cells on different days post irradiation and from non-treated control cells (NT). Primers for actin were used to verify integrity and quantity of cDNA; (B) Expression of REC8 protein in Namalwa cells, detected by Western blotting (antibody from Santa Cruz). The highest level of transcription was detected on day 3. Lower panel: for loading control PVDF membranes were re-blotted with mouse monoclonal anti-b-actin antibodies. (C) Transcription of SYCP1 and SYCP3 shown by RT-PCR of mRNA isolated from non treated Namalwa cells or on day 7 post irradiation showing very weak background expression. NB: In this experiment three times as much cDNA was used for the WI-L2-NS non treated samples as was used for the WI-L2-NS samples post irradiation. This was done in order to verify the increase in transcription after irradiation treatment. Although the amount of cDNA taken for PCR with the REC8 primers is much smaller on day 5 post irradiation (as judged by beta actin level) it still shows obviously higher level of REC8 expression.