Figure 1.

Induction of HLA-DR in human small intestinal enteroids. (A) Enteroids were stained with antibody to the MHC-II protein, HLA-DR, under standard culture conditions or following IFNg treatment (100 U/ml, 72 h) and imaged via fluorescence microscopy. White arrowheads indicate basolateral HLA-DR. (B) Median fluorescence intensity (MFI) of surface HLA-DR levels on enteroids treated for 24, 48, 72, or 96 h with 0–400 U/ml IFNg and analyzed by flow cytometry. Data are representative of two experiments. (C) RT-qPCR quantification of HLA-DR beta transcript in six enteroid lines, with GAPDH as a housekeeping gene. Error bars correspond to standard error of the mean. Statistical comparison of IFNg-treated and untreated enteroids performed by Wilcoxon Matched Pairs Signs Rank test. (D) Diagram of enteroid polarity reversal, whereby removal of extracellular matrix scaffold induces enteroids to adopt an apical-out morphology. (E) Apical-out and basolateral-out enteroids were treated with IFNg and imaged for HLA-DR or were pre-treated with IFNg prior to polarity reversal (F). Images in (E,F) are representative of three experiments. Scale bar = 20 μm. ^*p < 0.05.