Journal: Scientific Reports

Article Title: Discovery of a novel RORγ antagonist with skin-restricted exposure for topical treatment of mild to moderate psoriasis

doi: 10.1038/s41598-021-88492-1

Figure Lengend Snippet: Biochemical and cellular on-target activities of SHR168442. ( a ) Principle of the LanthaScreen TR-FRET human RORγ LBD coactivator assay. The terbium (Tb)-anti-GST antibody indirectly labels the RORγ LBD by binding to the GST tag of the protein. Binding of the compound to RORγ LBD causes a conformational change that results in a decrease in the affinity of the receptor for a labeled coactivator peptide, fluorescein-D22. ( b ) Dose–response curve for SHR168442 as an antagonist in LanthaScreen TR-FRET human RORγ LBD coactivator assay. ( c-d ) SHR168442 inhibits IL-17 secretion from human PBMCs ( c ) and mouse splenocytes ( d ). ( e–f ) Effects of SHR168442 on RORγ ( e ) and RORα ( f ) luciferase reporter expression. Ursolic acid and ATRA were used as positive control in the assays, respectively. RLU = relative light units. Non-linear regression analyses were performed and IC 50 values determined using GraphPad Prism software. Representative curves are shown. Average of the measurement and its standard deviation are shown.

Article Snippet: On day 5, compounds were added and then incubated for 72 h, and IL-17 in the supernatant was measured using IL-17A homodimer (mouse) AlphaLISA detection kit (Perkin Elmer) according to the manufacturer’s protocol.

Techniques: Binding Assay, Protein Binding, Labeling, Luciferase, Expressing, Positive Control, Software, Standard Deviation