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. 2010 Feb 16;38(11):3760–3770. doi: 10.1093/nar/gkq028

Figure 3.

Figure 3.

Hu proteins promote HuD exon 6 inclusion. (A) A schematic diagram of the HuD splicing reporter construct E6. The two potential splicing products are indicated. (B) E6 reporter was co-transfected into HeLa cells with pCDNA3.1 vector, TIAR, or different Hu protein isoforms. Splicing of the reporter was analyzed by RT-PCR on total RNA isolated from the transfected cells (top). The percentage of exon 6 inclusion is indicated below the RT-PCR gel. Expression of co-transfected proteins was analyzed by western blot assay (bottom). γ-Tubulin was used as a loading control. The molecular markers for DNA or protein size are indicated on the left of the gels. (C) Graphic representation of RT-PCR results for exon 6 inclusion of the E6 reporter shown in (B), with error bars indicating standard deviations. (D) F9 cells were transfected with pCDNA3.1 vector or HuC protein. Splicing of the endogenous HuD was analyzed by RT-PCR on total RNA isolated from the transfected cells (top). The percentage of HuD sv4 production is indicated below the RT-PCR gel. Expression of the over-expressed protein was analyzed by western blot (bottom). γ-tubulin was used as a loading control.