One glo lysis buffer

This assay was carried out as described [20 (link)]. HEK293 cells cotransfected with the plasmids of the hGLP-1R and luciferase reporter for cAMP (pGL4.29-Luc-CRE) or intracellular Ca²⁺ (pGL4.30-Luc-NFAT) or ERK phosphorylation (pGL4.33-Luc-SRE) were treated with increasing concentrations of agonist for 4h (cAMP and ERK) or 8h (Ca²⁺) at 37°C/5% CO₂. After the stimulation, an equal volume of ONE-Glo^™ lysis buffer containing luciferase substrate (Promega, Southampton, UK) was added to each well and luminescence measured using a plate reader in accordance with the manufacturer’s instructions.

Primary DRG neuronal cultures isolated as described above were transduced with pGreenFire1‐LXRE in LXRα Lentivector (System Biosciences, California) followed by respective group treatment 48 hrs after transduction. pGreenFire1‐LXREin LXRα co‐expresses a destabilized copepod GFP and luciferase from the LXR response elements and neighboring regions in the LXRα promoter paired with a mCMV promoter. After 24 hrs of treatment, cells were lysed in equal volume of ONE-GloLysis Buffer (Promega, Wisconsin) to that of culture medium. The luminescence was measured using CLARIOstar (BMG Labtech) and normalized to total protein.