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. 2022 Jul 1;298(8):102224. doi: 10.1016/j.jbc.2022.102224

Figure 5.

Figure 5

The U2AF2 UHM influences splicing of a prototypical minigene substrate.A, schematic diagram of the pyPY transcript. Alternatively spliced sites generate two differently sized products (py, 272 bp and PY, 131 bp). The unspliced pyPY RT-PCR product is 397 bp. B, representative RT-PCR of total RNA isolated from HEK 293T cells stably expressing the pyPY minigene and transfected with either wildtype (WT) or a double mutant U2AF2 (E394K/E397K). The RT-PCR products were resolved by agarose gel electrophoresis with ethidium bromide staining. C, quantification of pyPY RT-PCR products. The band intensities from three independent replicates were background corrected and averaged. The average ratios of the total spliced product (py + PY) to the unspliced pyPY and standard deviations are plotted for each sample. Primer sequences are listed in Table S3, and immunoblots are shown in Fig. S3. STD, 100 bp ladder molecular size markers. ∗p < 0.05.