Suppression of Peroxisome Proliferator-Activated Receptor -Coactivator-1 Normalizes the Glucolipotoxicity-Induced Decreased BETA2/NeuroD Gene Transcription and Improved Glucose Tolerance in Diabetic Rats

Supplemental Data

Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Glucolipotoxic conditions in pancreatic β-cells.
  A, INS-1 cells cultures at 5.5, 16.7 and 33.3 mM glucose for various times, and cell viability was measured using the MTT assay (*P < 0.05 and **P < 0.01). The results of the MTT assay from 10 independent experiments are given as mean ± standard error (SE). B, We performed AO/PI staining in freshly isolated rat islets. Isolated islets were incubated with or without 0.6 mM free fatty acids in the presence of a high concentration of glucose for 3 days to induce glucolipotoxicity. The glucolipotoxicity was associated with the gradual increase in β-cell death in a time-dependent manner compared with the low-glucose condition.
• Supplemental Figure 2 - Transfection efficiency of the adenovirus in INS-1 cells.
  INS-1 cells were transduced with 50 MOI of adenovirus for 2 h in serum-free DMEM. After infection with adenovirus, the INS-1 cells were incubated under low glucose condition for 3 days. A, Fluorescence in INS-1 cells was observed under a fluorescence microscope. B, The proportion of GFP⁺ cells was counted by flow cytometry on day 3. We confirmed that about 60% of totla INS-1 cells were GFP⁺ compared with the nonadenovirus-transfected INS-1 cells (P < 0.01). The flow cytometry data from the four independent experiments are summarized as mean ± standard error (SE).
• Supplemental Figure 3 - Trypan blue injection into the celiac artery.
  We used the superior pancreaticoduodenal artery, a branch of the celiac artery. First, we injected trypan blue to confirm the distribution of this artery by detecting the stained areas, which included the cranial portion of the pancreas, part of the stomach, and part of the duodenum, but not the liver. We concluded that the inferior pancreaticoduodenal artery kept free of a virus uniformly spread virus in the remaining head of the pancreas.