Fig. 3

cAMP-activated AVP promoter activity is modulated through Creb3l1. a-b AtT20 cells were transfected with 1 kb rat Avp promoter construct. At 24 h after transfection the cells were a treated with 10 μM FSK for 4 and 24 h or b pre-treated with 100nM DEX or DMSO for 2 h and followed by 4 h treatment of 10 μM FSK or DMSO and luciferase activity was measured (n = 3). c–e The Creb3l1-knockdown AtT20 cell line was produced in parallel with a control non-targeting shRNA cell line by transduction of lentivirus containing Creb3l1 or control non-targeting shRNA and further selected with puromycin. c The level of Creb3l1 knockdown was investigated by qPCR and immunoblotting in the AtT20 shRNA cell lines (n = 3). d The effect of FSK on Creb3l1 mRNA expression was examined by qPCR in control non-targeting and Creb3l1 knockdown cell lines (n = 3). e Luciferase assays were performed in control and Creb3l1-knockdown cell lines using 1 kb Avp promoter construct driving luciferase reporter gene (n = 3-4). Error bar + SEM; *, p < 0.05; **, p < 0.01; ***, p < 0.001 (Two way ANOVA for A, D and E, One way ANOVA for B and independent sample unpaired Student’s t-tests for C). FSK, forskolin; Con, Control; sh, shRNA; h, hour; ns, no significant difference