Journal: Scientific Reports

Article Title: Production and evaluation of parathyroid hormone receptor 1 ligands with intrinsic or assembled peroxidase domains

doi: 10.1038/s41598-017-13548-0

Figure Lengend Snippet: Luminescent detection of recombinant PTHR 1 s using the ligand PTH-APEX2. ( A ) Transfected HEK 293a (PTHR 1 or pcDNA3.1) grown in opaque white 24-well plates were stimulated with PTH-APEX2 for 30 minutes at 37 °C. Following this incubation the cells were incubated with the Western Lightning®Plus ECL substrate allowing the luminescent detection of PTHR 1 bound PTH-APEX2. ANOVA indicated that values were heterogenous (P < 10 −4 ). Tukey’s multiple comparison test was applied to compare pairs of values. For each vector, values vs. their respective controls: *P < 0.05; **P < 0.001; effect of adding an excess of PTH 1–34 vs. the enzymatic ligand alone: ‡ P < 0.001. Comparison of each experimental condition between vectors: † P < 0.001. ( B ) Endogenous receptors investigated in HOS cells. ANOVA indicated that values were heterogenous (P < 0.001). Each treatment vs. control value: *P < 0.01; **P < 0.001; effect of adding an excess of PTH 1–34 vs. the enzymatic ligand alone: not significant (Tukey’s multiple comparison test).

Article Snippet: HEK 293a cells transfected with the appropriate vector (either pcDNA3.1 or PTHR 1 ) or HOS cells were grown in white 24-well microplates with clear bottom (VisiPlate-24, PerkinElmer, Whaltham, MA, USA).

Techniques: Recombinant, Transfection, Incubation, Western Blot, Plasmid Preparation