Figure 2.

Ponalrestat rescues ethylene-induced short root growth downstream of EIN3. A, Western blotting for EIN3-GFP protein in EIN3-GFP/ein3 eil1. The seedlings were grown in the dark for 5 days on 1/2 MS medium with or without ponalrestat (5 μm) and/or ACC (10 μm). α-GFP antibody was used to detect the EIN3-GFP protein. Western blotting with actin antibody and membrane staining with Ponceau S were performed as the loading control. B, expression of 5xEBS:GUS in the roots of Col-0, ein3 eil1, and EIN3ox. The seedlings were grown in the dark for 3 days on 1/2 MS medium with or without ponalrestat (5 μm) and/or ACC (10 μm), followed by GUS staining. C, root lengths of etiolated seedlings of Col-0, eto1–2, ctr1–1, and EIN3ox grown on 1/2 MS medium supplemented with different concentrations of ponalrestat. Bars represent mean ± S.D. of at least 10 seedlings. Statistical differences between the groups were calculated with ANOVA (with Duncan's post hoc test). Bars with different letters are significantly different at p < 0.01. All the above experiments were repeated at least three times with similar results.