anti syntaxin 3  (Novus Biologicals)

Journal: Journal of pharmacological sciences

Article Title: SKF-10047, a prototype Sigma-1 receptor agonist, augmented the membrane trafficking and uptake activity of the serotonin transporter and its C-terminus-deleted mutant via a Sigma-1 receptor-independent mechanism.

doi: 10.1016/j.jphs.2018.11.005

Figure Lengend Snippet: Fig. 4. Effects of SKF-10047 on the protein expression of Syntaxin3 (STX3). Syntaxin3 consists of some isoforms, produced by alternative splicing (https://www.uniprot. org/uniprot/Q13277). According to the predictive molecular size of isoforms, the isoform A and isoform B, main isoforms of STX3 were considered to be detected in COS-7 cells (left). Treatment of COS-7 cells with SKF-10047 for 48 h significantly increased the protein levels of both STX3 isoforms in a dose-dependent manner. Statistical analysis was performed for isoform B (*P < 0.05, n ¼ 3, one-way ANOVA with Dunnett's post-test).

Article Snippet: An anti-Syntaxin3 rabbit polyclonal antibody was purchased from Alomone labs. (Jerusalem, Israel).

Techniques: Expressing, Produced, Alternative Splicing

Journal: Journal of pharmacological sciences

Article Title: SKF-10047, a prototype Sigma-1 receptor agonist, augmented the membrane trafficking and uptake activity of the serotonin transporter and its C-terminus-deleted mutant via a Sigma-1 receptor-independent mechanism.

doi: 10.1016/j.jphs.2018.11.005

Figure Lengend Snippet: Fig. 5. Influence of knockdown or overexpression of Syntaxin3 on the incremental effects of SKF-10047 on the SERTDCT uptake activity. We examined the effects of SKF-10047 on the SERTDCT uptake activity in STX3-knockdown or STX3-overexpressing COS-7 cells, and compared the increasing rate in these cells to that in control cells. A: Validation of STX3 knockdown by STX3 siRNA (upper) and overexpression by pMyc-DDK-STX3 (lower) in COS-7 cells. The transfection of STX3 siRNA #1 and #2 reduced the expression of STX3 isoform A, a functional isoform, by 53% and 56%, respectively (upper). In pMyc-DDK-STX3-transfected cells, prominent expression of Myc-DDK-STX3 was seen in addition to endogenous STX3, isoform A and B. B: Effects of SKF-10047 on the uptake activity of SERTDCT in control siRNA-transfected COS-7 cells and STX3 siRNA #2-transfected COS-7 cells. In both cells, 200 mM SKF-10047 significantly increased the uptake activity of SERTDCT (n ¼ 4, P < 0.001, unpaired t-test). The increase in SERTDCT uptake activity induced by 200 mM SKF-10047 was similar in control siRNA-transfected cells and STX3 siRNA-transfected cells (n ¼ 4, ns, unpaired t-test). C: Effects of SKF-10047 on the uptake activity of SERTDCT in mock-transfected COS-7 cells and Myc-DDK-STX3-expressing COS-7 cells. In both cells, 200 mM SKF-10047 significantly increased the uptake activity of SERTDCT (n ¼ 4, P < 0.001, unpaired t-test). The increase in SERTDCT uptake activity induced by 200 mM SKF-10047 in Myc-DDK-STX3-expressing cells was significantly higher than that in mock-transfected cells (n ¼ 4, P < 0.001, unpaired t-test).

Article Snippet: An anti-Syntaxin3 rabbit polyclonal antibody was purchased from Alomone labs. (Jerusalem, Israel).

Techniques: Knockdown, Over Expression, Activity Assay, Control, Biomarker Discovery, Transfection, Expressing, Functional Assay