Journal: bioRxiv
Article Title: FXR and BET signaling orchestrate to protect β cells
doi: 10.64898/2026.04.10.716420
Figure Lengend Snippet: a , Schematic of the differentiation strategy into HILOs. IHC of insulin Alexa-488 and DAPI of day 30 HILOs are shown. Illustration of the protocol for iPSC-derived human islet-like organoid (HILO) in vitro T1D and T2D models. T2D includes HILO with DOX-induced overexpression of human IAPP and TXNIP. The T1D HILO model is the co-incubation of HILO with peripheral blood mononuclear cells carrying the matched with HLA-A*01:01 HLA-B*08:01, HLA-C*07:01 HLA-DRB1*03:01 as the HILO. Scheme of partially HLA-matching was shown. b , Apoptosis rate in dispersed HILOs was measured by real-time imaging using Incucyte SX5. Dispersed HILOs were treated with DMSO, Fex (10 µM), JQ1 (500 nM), BD2i (1 µM), Fex (10 µM) + JQ1 (500 nM), or Fex (10 µM) + BD2i (1 µM), in the presence of IL-1β (10 ng/ml) and IFNγ (10 ng/ml). n = 3/each. Apoptosis was visualized by Caspase 3/7 green fluorescence. Bar graph shows green (apoptosis) cell count at 120 hours. c , Caspase 3/7 green (Green = Caspase 3/7 activity/Orange = human TXNIP/IAPPOE cells count) of T2D HILO model (DOX-inducible human TXNIP and human IAPP inducing) during 120 hours treatment of DMSO, Fex (10 µM), JQ1 (500 nM), BD2i (1 µM), Fex (10 µM) + JQ1 (500 nM), or Fex (10 µM) + BD2i (1 µM). n = 3/each. d , Apoptosis rate in dispersed HILOs was measured by real-time imaging using Incucyte SX5. Dispersed HILOs were co-cultured with human T1DPBMC and treated with Fex (10 µM), JQ1 (500 nM), BD1i (1 µM), or BD2i (1 µM) with or without IFNγ (10 ng/ml) for 120 hours. n = 3/each. e , Apoptosis rate in dispersed HILOs was measured by real-time imaging using Incucyte SX5. Dispersed HILOs were co-cultured with human T1DPBMC and treated with Fex (10 µM), JQ1 (500 nM), BD1i (1 µM), or BD2i (1 µM) with or without IFNγ (10 ng/ml) for 120 hours. Thap (Thapsigargin, ER stress inducer, 5 µM) were pretreated to enhance the expression of misfolded insulin prior to imaging. n = 3/each. Statistical analyses were performed using two-way ANOVA for the left panels of (b–e) and one-way ANOVA with Tukey’s multiple-comparison test for the right panels of (b–e). Error bars represent mean ± SEM.
Article Snippet: Caspase 3/7 cell apoptosis assays were performed using IncuCyte Caspase 3/7 green dye apoptosis reagent (Sartorius, 4440) and monitored using the IncuCyte SX5 live cell analysis instrument with according to the manufacturer’s protocol.
Techniques: Derivative Assay, In Vitro, Over Expression, Incubation, Imaging, Fluorescence, Cell Characterization, Activity Assay, Cell Culture, Expressing, Comparison
Elabscience Biotechnology is a verified supplier 
Elabscience Biotechnology manufactures this product 
