Prehybridized

The membranes were prehybridized with Hyb TM Plus buffer (Sigma) for 4 h followed by hybridization with the DNA probes overnight at 50°C. After washing the membranes twice with 5X SSC and once with 1X SSC and 0.1% SDS at 50°C, they were exposed for 2 days to radioactive sensitive screens, and scanned using a Thyphoon Trio scanner (AmershamBiosciences, New Jersey, USA).