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a , Endogenous PD-L1 expression highlighted in red in human islet cells (n= 3245) (β cells are outlined in red). b , Heatmap of top differentially expressed genes between PD-L1 + and PD-L1 - β cells. c , Immunohistochemistry showing overlap of <t>lentiviral-driven</t> PD-L1 expression and insulin promoter-driven GFP expression in wHILOs (scale bar, 100μm). d , Human PD-L1 and human insulin expression in wHILOs with and without lentiviral PD-L1 overexpression, as measured by qPCR (n=3). e , Transplantation of PD-L1 overexpressing wHILOs into the kidney capsule of STZ-induced diabetic mice. f , Blood glucose levels of C57BL6J mice treated with high dose streptozotocin (HD-STZ) prior to transplantation of wHILOs with and without PD-L1 overexpression (500 wHILOs into each kidney, total 1,000 wHILOs) (n=3) g , PD-L1 expression in human islet 12 hours after IFNγ stimulation (n=5). h , PD-L1 expression in wHILOs 12 hours after indicated IFNγ stimulation (n=3). Error bars represent ± SEM. *p<0.05, **p<0.01, ***p<0.001, one-tailed, student’s paired t test. Data were pooled from 3 independent samples (a) or representative of 3 independent experiments (c, d, g, h).
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a , Endogenous PD-L1 expression highlighted in red in human islet cells (n= 3245) (β cells are outlined in red). b , Heatmap of top differentially expressed genes between PD-L1 + and PD-L1 - β cells. c , Immunohistochemistry showing overlap of lentiviral-driven PD-L1 expression and insulin promoter-driven GFP expression in wHILOs (scale bar, 100μm). d , Human PD-L1 and human insulin expression in wHILOs with and without lentiviral PD-L1 overexpression, as measured by qPCR (n=3). e , Transplantation of PD-L1 overexpressing wHILOs into the kidney capsule of STZ-induced diabetic mice. f , Blood glucose levels of C57BL6J mice treated with high dose streptozotocin (HD-STZ) prior to transplantation of wHILOs with and without PD-L1 overexpression (500 wHILOs into each kidney, total 1,000 wHILOs) (n=3) g , PD-L1 expression in human islet 12 hours after IFNγ stimulation (n=5). h , PD-L1 expression in wHILOs 12 hours after indicated IFNγ stimulation (n=3). Error bars represent ± SEM. *p<0.05, **p<0.01, ***p<0.001, one-tailed, student’s paired t test. Data were pooled from 3 independent samples (a) or representative of 3 independent experiments (c, d, g, h).

Journal: Nature

Article Title: Immune evasive human islet-like organoids ameliorate diabetes

doi: 10.1038/s41586-020-2631-z

Figure Lengend Snippet: a , Endogenous PD-L1 expression highlighted in red in human islet cells (n= 3245) (β cells are outlined in red). b , Heatmap of top differentially expressed genes between PD-L1 + and PD-L1 - β cells. c , Immunohistochemistry showing overlap of lentiviral-driven PD-L1 expression and insulin promoter-driven GFP expression in wHILOs (scale bar, 100μm). d , Human PD-L1 and human insulin expression in wHILOs with and without lentiviral PD-L1 overexpression, as measured by qPCR (n=3). e , Transplantation of PD-L1 overexpressing wHILOs into the kidney capsule of STZ-induced diabetic mice. f , Blood glucose levels of C57BL6J mice treated with high dose streptozotocin (HD-STZ) prior to transplantation of wHILOs with and without PD-L1 overexpression (500 wHILOs into each kidney, total 1,000 wHILOs) (n=3) g , PD-L1 expression in human islet 12 hours after IFNγ stimulation (n=5). h , PD-L1 expression in wHILOs 12 hours after indicated IFNγ stimulation (n=3). Error bars represent ± SEM. *p<0.05, **p<0.01, ***p<0.001, one-tailed, student’s paired t test. Data were pooled from 3 independent samples (a) or representative of 3 independent experiments (c, d, g, h).

Article Snippet: Lentiviruses were produced using second- or third-generation lentiviral systems in HEK293LTV cell line (Cell Biolabs).

Techniques: Expressing, Immunohistochemistry, Over Expression, Transplantation Assay, One-tailed Test