Journal: International Journal of Molecular Sciences

Article Title: Crosstalk between β2- and α2-Adrenergic Receptors in the Regulation of B16F10 Melanoma Cell Proliferation

doi: 10.3390/ijms23094634

Figure Lengend Snippet: Effects of β-AR and a-AR ligands on cAMP production in B16F10 cells: ( A ) Representative image of cAMP measurements in real time using a GloSensor cAMP biosensor (bas, baseline). ( B ) Integrated cAMP responses computed as % of integrated forskolin response from tracings obtained in B16F10 cells stably expressing GloSensor-22F probe. Measurements were obtained in the absence or the presence of the β-AR agonist isoproterenol (ISO 1 μM), the antagonists propranolol and ICI 118,551 (PRO 10 μM, ICI 118,551 10 μM), isoproterenol plus either propranolol or ICI 118,551 (ISO 1 μM + PRO 10 μM or ICI118,551 10 μM), and the α2 agonists clonidine (CLO 1 μM) and ST91 (ST-91 1 μM). Data of three independent experiments are reported. * p < 0.05. ( C ) Concentration–response curves for the ligand-induced enhancement of cAMP production. Epinephrine (EPI), isoproterenol (ISO), norepinephrine (NE). EC 50 for ISO, EPI and NE was 4.6 nM, 45 nM and 284 nM, respectively.

Article Snippet: The luciferase-based intracellular cAMP probe GloSensor 22 F was purchased from Promega (Madison, WI, USA) and subcloned into puromycin resistance retroviral expression vector pQCXIP (Clontech, Mountain View, CA, USA).

Techniques: Stable Transfection, Expressing, Concentration Assay

Journal: The FASEB Journal

Article Title: Regionally selective cardiovascular responses to adenosine A 2A and A 2B receptor activation

doi: 10.1096/fj.202101945R

Figure Lengend Snippet: Glosensor cyclic AMP responses to CGS 21680 and BAY 60‐6583 in HEK293G cells. (A) Concentration‐response curves for agonist‐stimulated cAMP Glosensor luminescence responses in HEK293G cells in response to CGS 21680 and BAY 60‐6583. (B) The effect of simultaneous addition of 100 µM BAY 60‐6583 on the concentration‐response curve to CGS 21680, following 2 h pretreatment with PSB 603 (0.1 µM). Also shown is the effect of simultaneous addition of 10 µM SCH 58261 on the responses to CGS 21680 in the presence of PSB 603 (0.1 µM). (C, D) The effect of 100 µM BAY 60‐6583 on the response to 10 µM formoterol in the (C) absence and (D) presence of 0.1 µM PSB 603. (E) The effect of simultaneous addition of 10 µM SCH 58261 on the response to 10 µM formoterol. Data represent mean ± SEM of the peak luminescence response of 5 separate experiments, each carried out in triplicate. Data are expressed as a percentage of the peak luminescence response obtained with 100 µM NECA measured in the same experiment. ** p < .05 (unpaired t ‐test)

Article Snippet: A clonal HEK 293 cell line stably expressing the cAMP GloSensor (20F) biosensor (HEK293G) , was obtained from Promega (Madison, WI, USA).

Techniques: Concentration Assay