Journal: Scientific reports

Article Title: Selenocysteine insertion sequence binding protein 2 (Sbp2) in the sex-specific regulation of selenoprotein gene expression in mouse pancreatic islets.

doi: 10.1038/s41598-020-75595-4

Figure Lengend Snippet: Figure 3. Selenoprotein gene expression analysis in Sbp2-deficient male and female mouse islets. Quantitative real time-PCR analysis (qPCR) of selenoprotein mRNA in male (a) and female (b) Sbp2 βCKO islets compared with that of control mouse islets (Sbp2 fl/fl). Individual selenoprotein mRNA expression is displayed compared relative to the expression of Gpx3. Expression was normalized to the housekeeping gene Hprt. Genes susceptible to NMD are marked with the following symbol: ‡. Representative immunoblots showing Gpx3, Selenop, Dio1, and Txnrd2 protein expression in male (c) or female (d) islets of Sbp2 βCKO and control (Sbp2 fl/fl) mice are shown; lanes 1 and 2 are representative islets from control mice (Sbp2 fl/fl), and lanes 3 and 4 are representative islets from Sbp2 βCKO islets; β-actin was used as the protein loading control. Corresponding quantification expressed in arbitrary units is shown at the right normalized to β-actin expression. (See also Supplementary Information 1.) Gene expression of the Selenop receptors Lrp1 and Lrp8 in male and female Sbp2 βCKO islets compared with that of control mouse islets (Sbp2 fl/fl) is shown in (e). Between group differences were analyzed by unpaired, two-tailed Student’s t test. N = 6 mice per group; p < 0.05 was considered statistically significant with individual p values as shown.

Article Snippet: Primary antibodies against Sbp2, Gpx3, Dio1, and Txnrd2 (all from Proteintech) as well as Selenop (Santa Cruz) were used to quantify select selenoprotein expression by immunoblotting after separation by SDS-PAGE; β-actin (Cell Signaling) was used as the internal protein loading control.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Control, Expressing, Western Blot, Two Tailed Test