Figure 2.

Tissue distribution of ELL2 and ELL mRNAs. A human multiple tissue northern blot (MTN1, CLONTECH) was probed sequentially with PCR-generated ELL2- and ELL-specific probes chosen from a region of sequence that was most divergent between the two genes. The ELL-specific probe contained sequences encoding amino acids 317–621, and the ELL2-specific probe contained sequences encoding amino acids 327–474. As a loading control, the same blot was probed with a probe specific for glyceraldehyde-3-phosphate dehydrogenase (G3PDH) mRNA. Probes were labeled with [α-³²P]dCTP by random priming performed according to the manufacturer’s instructions (Rediprime kit, Amersham). The blot was prehybridized in 10 ml of Hybrisol I solution (Oncor) for 3 h at 42°C. Probe DNA was denatured and added to hybridization solution at 10⁶ cpm/ml of solution. Hybridization was carried out at 42°C overnight. The blot was washed 10 min in 2× standard saline citrate (SSC)/0.1% SDS at room temperature, 15 min in 0.2× SSC/0.1% SDS at 45°C, 10 min in 0.1× SSC/0.1% SDS at 55°C, and then exposed to film (Hyperfilm-MP, Amersham) overnight at −80°C.