A. RA enhances the stimulatory effect of Acinus on the splicing RARE-tubg1E8-E9 pre-mRNA
in a dose-dependent manner. B. The synergism of RA and Acinus in promoting the splicing of
RARE-tubg1E8-E9 pre-mRNA is time-dependent. In Panels A and B, 293A cells were transfected
with pRARE-tubg1E8-E9 splicing reporter DNA, RARβ expression vector DNA, and either
V5-Acinus-S’ (S’) expression vector DNA or V5 empty vector DNA (V).
Twenty-four hr after transfection cells were treated with ethanol or RA at the indicated
concentrations (A) or 10−6 M (B) for 24 hr (A) or times ranging from 0
to 24 hr (B). Total RNA was prepared and used to analyze splicing as described in the
legend to Figure 1. Values represent mean ± SD
from 3 independent experiments. V, V5 empty vector; S’, V5-Acinus-S’; E,
ethanol; R, retinoic acid. * p < 0.05, ** p
< 0.01, *** p < 0.001, unpaired t
test.