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. 2019 Nov 1;4(21):e131028. doi: 10.1172/jci.insight.131028

Figure 8. A20 expression inhibits human islet inflammation.

Figure 8

(A) Human islets transduced with an adenovirus encoding for GFP (rAd.GFP) at multiplicity of infections indicated or left noninfected (NI). Twenty-four hours after transduction, islets were digested to single cells, and the percent of GFP+ cells was determined by flow cytometry. Each dot per column represents an independent human donor. (B) Donor human islet preparations were transduced with rAd.GFP at a MOI of 10:1 or left noninfected (NI). Forty-eight hours after transduction, islets were fixed for immunofluorescence analysis. Assessed proteins are indicated below each panel. Scale bar: 50 μm. (C and D) Donor human islets transduced with rAd.GFP or A20 at a MOI of 10:1. Forty-eight hours after transduction, cells were lysed and assessed for (C) A20 RNA expression and (D) protein levels. (E) Function of infected islets were assessed in a glucose-stimulated secretion assay. Stimulation index indicates the amount of insulin in supernatant in high glucose/low glucose. (F) GFP- and A20-transduced donor islets stimulated with TNF for the indicated times and expression of inflammatory genes assessed. Data in A–F are cumulative from 3 independent human donor islet preparations. Error bars ± SEM and statistical significance determined by 2-way ANOVA with Sidak’s multiple comparisons post hoc test; * P < 0.05; ****P < 0.0001.