Fig. 1. Expression patterns of the Dd-ef2h mRNA and the Dd-EF2H protein during early development in Ax-2 cells and various transformed cells. Cells were harvested at the exponential growth phase, washed twice in BSS and shaken for the indicated times (hours) at 22°C. Total RNAs were prepared as described (Nellen et al., 1987). Northern hybridization was performed using the RI (Amersham), as previously described (Hirose et al., 2000). As a probe for detection of the Dd-ef2h mRNA, a PCR-product (3 kb) obtained by amplification of the cDNA clone SLE406 with the full-length of Dd-ef2h using M13-20 and M13R primers was used. Western blotting was performed as described in Materials and Methods. The expression patterns in ef-2^AS cells underexpressing the Dd-ef2h mRNA (A), Dd-ef2 null cells produced by homologous recombination (B), and ef-2^OE cells overexpressing the Dd-ef2h mRNA (C) are presented in comparison with those in parental Ax-2 cells. In the lower panel in B, the amount of actin in each lane (stained with CBB) is shown.