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. Author manuscript; available in PMC: 2016 Apr 1.
Published in final edited form as: J Cell Physiol. 2015 Apr;230(4):791–801. doi: 10.1002/jcp.24804

Figure 3. The synergistic effect of RA and Acinus on the splicing of a RA-responsive reporter minigene containing a weak 5′ splice site is RA dose-dependent and time-dependent.

Figure 3

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.