Forskolin (Coleonol) is a natural product, an adenylate cyclase activator (EC50=0.5 μM). Forskolin increases cAMP levels, activates PXR and FXR, and induces autophagy. Forskolin produces positive inotropic effects in the heart, and has platelet anticoagulant and antihypertensive effects.

adenylyl cyclase (type I):0.5 μM (EC50), MCF-7 cells:63.3 μM, Adenylyl cyclase:0.5 μM (cell free)

METHODS: Rat adrenal medullary chromosomal tumor cells PC12 were treated with Forskolin (0.01-10 µM) for 3-48 h. The growth inhibition of the cells was detected by MTT.
RESULTS: The cell viability decreased rapidly after treatment with 10 µM Forskolin (88.4% after 6 h and 60.5% after 48 h). [1]
METHODS: Human myeloma cells U266, H929, INA-6, RPMI 8226 and OPM-2 were treated with Forskolin (1-100 µM) for 72 h. Cell death was detected by Flow Cytometry.
RESULTS: Forskolin dose-dependently induced cell death in human myeloma cells, with U266, OPM-2 and INA-6 being more sensitive than H929 and RPMI 8226 cells. [2]
METHODS: Human IL-2-dependent leukemia cells Kit 225 and human leukemia cells MT-2 were treated with Forskolin (1-100 μM) for 20 min, and cAMP concentration was measured by ELISA.
RESULTS: Forskolin induced an up-regulation of cAMP levels, which reached a maximum level between 50-100 μm. [3]

METHODS: To assay anti-tumor activity in vivo, Forskolin (4-5 mg/kg in PBS/DMSO solution (15:1)) was injected intraperitoneally into BALB/c nude mice bearing murine multiple myeloma tumor MOPC315, and was administered on the 2nd/4th/6th day after tumor cell injection.
RESULTS: All mice eventually developed tumors, but Forskolin significantly delayed tumor growth in vivo. Compounds that increase cAMP may have therapeutic potential in the treatment of multiple myeloma. [4]
METHODS: To investigate the effect of Forskolin on retinal inflammation under diabetic conditions, Forskolin (50 mg/kg) was administered by gavage once weekly for twelve weeks to C57BL/6 mice in the STZ-induced diabetes model.
RESULTS: Retinal glucose concentrations were increased in both diabetic control and Forskolin-treated groups compared to normal controls, but due to down-regulation of glucose transporter protein 1 expression, the Forskolin-treated group was only about 68.06% of the diabetic control group. ICAM-1 and TNF-α expression was up-regulated in the Forskolin-treated and diabetic control groups compared to the normal control group, but the expression levels of these two inflammatory factors in the Forskolin-treated group were 68.75% and 75.37% of those in the diabetic control group, respectively. [5]

For Jak3 kinase assays, Fsk-treated MT-2 cells were lysed, clarified, and immunoprecipitated using Jak3 antibody as described above. Kinase reactions were carried out as described previously at 30 °C for 20 min. For PKA kinase assays, untreated MT-2 cells were lysed, and Jak3 was immunoprecipitated and bound to PAS beads as described previously. Immunoprecipitated Jak3 was washed with kinase buffer (50 mM Hepes-NaOH (pH 7.4), 10 mM MgCl2, 0.5 mM EGTA, 0.5 mM DTT, 20 μg/ml aprotinin, 10 μg/ml leupeptin, 1 μg/ml pepstatin A) and incubated with 200 μM ATP and purified protein kinase A catalytic subunit (PKAc) as indicated in the figure legends. Kinase reactions were carried out at 32 °C for 30 min followed by vigorous washing of the beads with cold kinase wash buffer as described previously. For [γ-32P]ATP radiolabeled kinase assays using recombinant Jak3, Hek293 cells were transfected with wild type (WT) Jak3 or kinase-dead Jak3 K855A using Lipofectamine 2000 according to the manufacturer's instructions. Cells were lysed and immunoprecipitated with Jak3 antibody. Jak3-bound PAS beads were washed three times in cold lysis buffer followed by kinase buffer. Kinase reactions were initiated by adding 10 μCi [γ-32P]ATP, 10 μm unlabeled ATP, and 1 μg of purified PKAc to Jak3-bound PAS bead reaction mixtures. Kinase reactions were performed at 32 °C for 30 min. Jak3-bound PAS beads were washed three times in radioimmunoassay buffer (10 mM Tris-HCl, pH 7.4, 75 mM NaCl, 20 mM EDTA, 10 mM EGTA, 20 mM Na4P2O7, 50 mM NaF, 20 mM 2-glycerolphosphate, 1 mM p-nitrophenyl phosphate, 0.1% Triton X-100) and one time in kinase wash buffer. The reactions were stopped by adding 2× SDS-PAGE sample buffer followed by SDS-PAGE. Coomassie stainable Jak3 bands were excised from the PVDF membrane and subjected to phosphoamino acid analysis [2].

Kit 225 or MT-

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