Deregulated PDX-1 expression in Ran-targeted cells.

<p><i>A</i>, Pancreas sections from non-TG, Ran-WT, Ran-G19V or Ran-T24N transgenic mice were stained with an antibody to insulin or PDX-1, by immunohistochemistry. <i>B</i>, The number of insulin- or PDX-1-stained cells was quantified by morphometry in the indicated surface area. Insulin⁺ cells, non-TG (n = 2), 28.2±0.4; Ran-WT (asymptomatic, n = 3), 29.8±1.2; Ran-WT (diabetic, n = 3), 6.2±0.6, ***, p<0.0001; PDX-1⁺ cells, Non-TG (n = 2), 34.2±4.1; Ran-WT (asymptomatic, n = 3), 37.5±2.3; Ran-WT (diabetic, n = 3), 22.8±1.8, **, p = 0.007. <i>C</i>, Islets from 2 mo-old non-TG or a Ran-WT transgenic mouse were analyzed by Western blotting. <i>D</i>, Pancreatic islets isolated from non-TG mice were transduced <u>ex vivo</u> with control pAd-GFP or pAd-GFP-Ran-WT, pAd-GFP-Ran-G19V or pAd-GFP-T24N and analyzed after 48 h by fluorescence microscopy for GFP expression (<i>left</i>), or Western blotting (<i>right</i>). *, non-specific. <i>E</i>, INS-1 cells were left untreated (INS-1) or transduced with control lentivirus (pLKO) or lentivirus encoding Ran-directed shRNA (Ran, 74V1), and analyzed by Western blotting. <i>F</i>, Parental INS-1 cells or four independent clones of INS-1 cells stably transduced with Ran-directed shRNA (77V-2, 74V-1, 75V-1, 75V-2) were analyzed for changes in insulin release in the supernatant. Representative experiment out of at least two independent determinations.