Figure 1
From: Establishment and phenotyping of disease model cells created by cell-resealing technique
Establishment of healthy and diabetic model H4IIEC3 cells using the cell-resealing technique. (a) Resealed cells labeled with propidium iodide (PI, red) and dextran conjugated with fluorescein (dextran, green). Bar = 10 µm. (b) Scheme for the establishment and analysis of healthy or diabetic model H4IIEC3 cells. (c) Resealed H4IIEC3 cells that were introduced with WT liver cytosol at the protein concentration of 0.0, 0.5, 1.5, or 3.0 mg/ml were incubated with DMEM(-FBS) for 1 hours, and further with or without insulin for 1hr. The relative expressions of PCK1 upon insulin treatment to the one without insulin are analyzed and the means and standard deviations from four independent experiments are shown in the graph. (d) and (e) The relative expression of PCK1 (d) or G6PC (e) in the presence or absence of insulin treatment in HWT (WT) and HDb (Db) cells. The expression level of PCK1 or G6PC in WT cells was set to 100%. The means and standard deviations from three independent experiments are shown in the graph. **p = 0.08389 × 10−6 in (d) and **p = 0.0092 in (e). (f) HWT and HDb cells were incubated in glucose-free medium with (WT + ins or Db + ins) or without (WT or Db) insulin at 37 °C for 3 hrs. The amount of glucose in medium was quantified by glucose assay. The amount of glucose of WT cells was set to 100%. The means and standard deviations from three independent experiments are shown in the graph. **p = 0.00284. (g) HWT or HDb cells were incubated with DMEM(-FBS) for 1, 6, 12, and 24 hours, and further with or without insulin for 1hr. The expression level of PCK1 was analyzed by real-time PCR, and the relative expression of PCK1 upon insulin treatment to the one without insulin at each time points are analyzed in HWT cells (○) or in HDb cells (●) and the means and standard deviations from three independent experiments are shown in the graph.
