Fig. 4.

Pretreatment with GPR107 siRNA prevents neuronostatin-induced PKA phosphorylation in mouse pancreatic cell line. Mouse pancreatic αTC1-9 clone cells were incubated for 48 h, pretreated with GPR107, GPR56, or eGFP siRNA for 28 h and then changed to low glucose media overnight (∼16 h). Neuronostatin or saline vehicle were added to the media for 40 min. Cells were lysed, protein was isolated and quantified, and 100 pg/sample was loaded on to a gradient gel and transferred to a PVDF membrane. The membrane was probed for phosphorylated PKAc (Thr-197), stripped, and reprobed for total PKAc (α). A: representative Western blot. Note that the order of treatment group (±) differs in vehicle-pretreated cells from other siRNA groups. B: average densitometry for separate experiments performed in duplicate. Significance to vehicle-treated controls is indicated by * (P < 0.05), as determined by a Mann-Whitney U-nonparametric test for transformed data.