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. 2008 Sep;57(9):2413–2420. doi: 10.2337/db07-1323

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Copyright © 2008, American Diabetes Association

Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details.

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FIG. 5. — Prevention of HES-1 upregulation by shRNA reduces β-cell dedifferentiation. A: qPCR analysis of RNA extracted from islet cells following infection with HES1 shRNA TRCN18993 or nontarget virus. RQ indicates relative quantification compared with P0. Data are means ± SD (n = 3 donors). The changes in all three genes in cells infected with HES1 shRNA, compared with cells infected with nontarget virus, were significant (P < 0.004). ▪, P0; □, nontarget;, HES1 shRNA. B: Left: Incidence of insulin-positive cells among cultured islet cells following infection with HES1 shRNA TRCN18993 or nontarget virus. Data are means ± SD (n = 3 donors; >1,000 cells counted in culture from each donor; P = 0.016). Right: Incidence of insulin-positive cells among eGFP⁺ β-cells from two representative donors following infection with HES1 shRNA or nontarget virus. Data are based on >1,000 cells counted in culture from each donor. Significant differences compared with nontarget are indicated by stars. □, G13; ▪, I10. C: Immunofluorescence analysis of insulin in eGFP⁺ cells following infection with HES1 shRNA TRCN18993 or nontarget virus. Bar = 100 μm. All the analyses were done 14 days following viral infection. (Please see http://dx.doi.org/10.2337/db07-1323 for a high-quality digital representation of this figure.)

Inline graphic — Prevention of HES-1 upregulation by shRNA reduces β-cell dedifferentiation. A: qPCR analysis of RNA extracted from islet cells following infection with HES1 shRNA TRCN18993 or nontarget virus. RQ indicates relative quantification compared with P0. Data are means ± SD (n = 3 donors). The changes in all three genes in cells infected with HES1 shRNA, compared with cells infected with nontarget virus, were significant (P < 0.004). ▪, P0; □, nontarget;, HES1 shRNA. B: Left: Incidence of insulin-positive cells among cultured islet cells following infection with HES1 shRNA TRCN18993 or nontarget virus. Data are means ± SD (n = 3 donors; >1,000 cells counted in culture from each donor; P = 0.016). Right: Incidence of insulin-positive cells among eGFP⁺ β-cells from two representative donors following infection with HES1 shRNA or nontarget virus. Data are based on >1,000 cells counted in culture from each donor. Significant differences compared with nontarget are indicated by stars. □, G13; ▪, I10. C: Immunofluorescence analysis of insulin in eGFP⁺ cells following infection with HES1 shRNA TRCN18993 or nontarget virus. Bar = 100 μm. All the analyses were done 14 days following viral infection. (Please see http://dx.doi.org/10.2337/db07-1323 for a high-quality digital representation of this figure.)