Figure 4. .

Validation of PNN-regulated splicing events in HCET cells. (A, B) Validation of alternatively spliced genes upon PNN knockdown was achieved by semi-quantitative RT-PCR assays with multiple primer pairs for FOXJ3 and FAM50A. Diagrams show exon structure around alternative region and the location of primers (colored arrows) used for RT-PCR validation. Reduction of PNN led to the increased retention of introns 11 and 9 in their entire length on FOXJ3 and FAM50A, respectively. Inclusion percentage of retained introns is shown under the corresponding lanes. (C–F) Examples of validated PNN-regulated splicing events are presented. While alternative splicing changes of PSENEN, NCSTN, and ECT2 are PNN-silenced events, inclusion of intron 9 of GLT8D1 is a PNN-enhanced event. All PCR products shown were verified by sequencing.