Figure 2.

Differential detection of the predominant mRNA variant forms for SUMO1, SUMO2, and SUMO3 by RT-qPCR analyses. (a) Schematic of the primer pairs used to specifically amplify each mRNA variant for SUMO1, SUMO2, and SUMO3. Most of the primers correspond to exon-exon junctions, thus providing selectivity for specific splicing events; the precise location of the exon junctions in the primers, their sequence, and orientation are indicated. Reverse primers are written upside down. The length of the PCR product amplified by every primer pair is shown under the name of the variant (left, in base pairs [bp]). (A)₂₅₀: Polyadenyl tail. (b) RT-qPCR products obtained for each mRNA variant form using the primer pairs represented in A. All products obtained were sequenced to confirm their identity and the specificity of the reaction. (c) Sequence analysis of the RT-qPCR product for SUMO2V2 showing the alternative splicing event that results in the juxtaposition of Exon 2 and Exon 4, as expected for SUMO2V2. Similar analyses confirming all predicted Exon-Exon junctions were performed for all RT-qPCR products.