Induction of Reg gene by IL-6/dexamethasone/PARP
inhibitors. RINm5F β cells were treated with IL-1β, TNFα, IFNγ,
IL-6, dexamethasone (Dx), nicotinamide (NA), 3-aminobenzamide (3AB),
Reg protein, and anti-rat Reg monoclonal antibody (17) (αReg). Values
represent mean ± SEM of at least seven experiments. Statistical
analyses were performed using Student's t test.
(A) RT-PCR analysis. Expression of Reg
and of the control glyceraldehyde-3-phosphate dehydrogenase (G3PDH)
gene were analyzed by RT-PCR. (B) WST-1 assay. Reg (1.5
μg/ml) was added to the medium. WST-1 cleavages were significantly
increased by the incubation with IL-6/Dx, IL-6/Dx/NA,
IL-6/Dx/3AB, and Reg (P < 0.001).
(C) BrdUrd incorporation. Reg (1.5 μg/ml) and αReg
(15 μg/ml) were added to the medium. (D) Immunoblot
analysis. Secreted Reg protein was analyzed by immunoblot of the
culture medium of the RINm5F cells.