FIG 7.

CDP138 is predominantly expressed in adrenal medulla and required for both basal and stressed-induced adrenaline release from adrenal glands. (A) Plasma adrenaline levels were measured by ELISA from 20-week-old mice fed with NCD (n = 4 in WT group and n = 6 in KO group) (left), 20-week-old mice fed with an HFD for 8 weeks (n = 8 in WT group and n = 7 in KO group) (middle), and 12-week-old mice fed with NCD plus cold challenge (CC) for 2 h (n = 6 in WT group and n = 7 in KO group) (right). (B) Adrenaline levels in total lysates of adrenal glands were measured by ELISA from 20-week-old mice fed with HFD for 8 weeks and 12-week-old female mice fed with NCD plus cold challenge for 2 h. (C) Immunofluorescence of CDP138 with adrenal gland tissue sections from 20-week-old WT and KO mice. Scale bars, 100 μm. (D) Adrenal gland tissue lysates from 20-week-old female WT and KO mice were immunoblotted with anti-tyrosine hydroxylase (TH) and anti-CDP138 antibodies. (E) Isolated adrenal gland of 12-week-old female WT and KO mice were minced and washed three times before incubation in 1 ml of enriched Dulbecco's modified Eagle medium for 30 min at 37°C. Conditional medium (200 μl) was taken for the basal adrenaline release measurement, and then the minced tissues were treated with 100 μM nicotine for 5 min to stimulate adrenaline release (n = 9 in each group). All data are presented as means ± SEM. P < 0.05 (*) and P < 0.01 (**) for WT versus KO mice; #, P < 0.01 for basal versus nicotine-stimulated adrenal medulla cells.