C0003 04

The following adenoviruses were purchased: Ad-GFP-m-SIK1 (ADV-271991, Vector Biolabs) and Ad-CMV-GFP (1060, Vector Biolabs) as a control. Dissociated mouse islets were transduced with adenoviruses for 2 h in RPMI 1640 medium (21875034, Thermo Fisher Scientific) without FBS at an MOI of 50. The wells were then washed, and RPMI 1640 (21875034, Thermo Fisher Scientific) containing 10% FBS and 15 mM glucose was added to the wells. After 4 h, cells were treated with HG (0.1, 0.05 or 0.025 μM) or with DMSO (1%) in the presence of EdU (10 μM). After 4 d, cells were either fixed for immunofluorescence studies or lysed to assess mRNA expression. Sik1 and Hprt (control) primer sequences are shown in Supplementary Table 8. For experiments with MIN6, cells were transduced with adenoviruses for 4 h in DMEM-based medium (C0003–04, AddexBio) without FBS at an MOI of 50. The wells were then washed, and DMEM-based medium (C0003–04, Addexbio) containing 15% FBS and 0.055 mM β-mercaptoethanol was added to the wells. After 4 d, cells were treated with HG (1 μM) or DMSO (1%) for 6 h. For the experiments presented in Fig. 7, islet cells from Atf6 floxed mice were transduced with Ad-LacZ (1080, Vector Biolabs) or Ad-Cre (1045, Vector Biolabs) at an MOI of 10 for 72 h in RPMI 1640 medium containing 10% FBS and 15 mM glucose.