Fig. 2.

Verification and quantification of the full-length and truncated mouse sst5 variants in mouse tissues. a Schematic representation of the full-length sst5 and novel truncated sst5 variant transcripts. CDS are shown by boxes (grey boxes CDS of sst5 or CDS_1 of each truncated sst5 variant; white boxes CDS_2 of each truncated sst5 variant). Arrow heads are used to indicate the relative positions of primers sets used in quantitative real-time RT-PCR (qrtRT-PCR; closed arrows indicated primers within a CDS, while open arrows indicate primers spanning CDS_1 and CDS_2). b Representative agarose gel showing PCR products amplified by qrtRT-PCR using mouse pituitary as template. All products (sst5 variants and cyclophilin used as a housekeeping gene) were size separated on an agarose gel containing EtBr, column-purified and sequenced to confirm target specificity. PCR product of the sst5TMD4 variant could not be amplified if pituitary cDNA was used as a template, indicating that this variant was not present in this tissue; however, the sst5TMD4 variant transcript was detected in other tissues (i.e., fat; Supplemental Figure 3). c Melting curves of qrtRT-PCR products with different T_m of dissociation (80.9°C for full-length sst5, 79.4°C for sst5TMD2 and 86.8°C for sst5TMD1) generated from the same mouse pituitary sample. d Absolute cDNA copy number/0.1 μg total RNA of full-length and truncated sst5 variant transcripts in the pituitary, hypothalamus, cortex and cerebellum of male C57Bl/6 mice, as determined by qrtRT-PCR. Values represent means ± SEM (n = 5 mice)