Figure 3
From: The CB1 cannabinoid receptor signals striatal neuroprotection via a PI3K/Akt/mTORC1/BDNF pathway
Multiple transcription factors are involved in the CB1 receptor-mediated induction of BDNF promoter IV. (a) STHdhQ7/Q7 cells were transfected with a construct harboring a WT 0.5-kb human BDNF promoter IV fused to the luciferase reporter gene or with the same construct containing mutations (m) in the indicated response elements (RE). Cells were subsequently incubated for 15 h with vehicle, 0.5-μM THC or 10-nM HU-210. Promoter activity relative to vehicle incubations is shown (n=4–6 experiments). (b) STHdhQ7/Q7 cells were transfected with a non-targeted siRNA or siRNAs directed against the indicated transcription factors. After 24 h, they were transfected with the aforementioned WT BDNF promoter IV reporter construct, and, after an additional 24-h period, cells were incubated with vehicle, 0.5-μM THC or 10-nM HU-210 for 15 h. Left: promoter activity relative to vehicle incubations; right: knock-down (KD) efficacy of the siRNAs directed against the corresponding transcription factor (siTF; n=4–6 experiments). Data were analyzed using unpaired Student’s t-test. *P<0.05 and **P<0.01 from the corresponding vehicle-treated cells or siControl-transfected cells (b, left). (c) STHdhQ7/Q7 cells were incubated for 30 min with vehicle, 0.5-μM THC or 10-nM HU-210, lysed and used for western blot analyses. Quantification of mean optical density (O.D.) values of pCREB relative to those of total CREB as well as a representative blot are shown (n=3–4 experiments)
