Figure 5.

Effect of cycloheximide treatment on JA-responsive genes. A, Twelve-day-old wild-type seedlings grown in liquid medium were treated with either a mock control (0.2% DMSO), 50 μm MeJA (MJ), 50 μm cycloheximide (CHX), or a combination of MeJA and CHX (MJ + CHX) as described in “Materials and Methods”. Whole seedlings were collected for RNA extraction at the indicated times (h) after treatment. Five micrograms of total RNA was loaded in each lane and blots were hybridized to the indicated probes (see text). ACT8 was used as a loading control. LOX3- and LOX4-probed blots were exposed to film for 14 h, whereas all other blots were autoradiographed for 3 h. B, Effect of coi1-1 on CHX-induced expression of JA-responsive genes. CHX treatment and RNA gel-blot analysis were performed as described above. All blots were autoradiographed for 4 h.