Journal: bioRxiv

Article Title: Protective IFIH1 variant reduces immune-mediated islet stress and dysfunction in a type 1 diabetes genetic background

doi: 10.64898/2025.12.30.697107

Figure Lengend Snippet: (A) , Workflow schematic for treatment processing of SC-islets. (B) , Harmony UMAP of cell populations identified following scRNA-seq data integration. (C) , Heatmaps depicting the z-score of genetic marker expression per cluster for each time point. (D) , Stacked bar plots depicting the proportion of cells found in SC-islets derived from each MDA5 variant and treated with various stressors for 48 h. (E-J) , Volcano plots showing transcriptional differences when comparing (E) MDA5 946T Ctrl vs IFNα 48 h (total variables = 1920), (F) MDA5 946T Ctrl vs poly(I:C) 48 h (total variables = 769), (G) MDA5 946T Ctrl vs CVB3 48 h (total variables = 3326), (H) MDA5 627* Ctrl vs IFNα 48 h (total variables = 1700), (I) MDA5 627* Ctrl vs poly(I:C) 48 h (total variables = 1083), and (J) MDA5 627* Ctrl vs CVB3 48 h (total variables = 1815).

Article Snippet: Whole SC-islets were then treated with either endotoxin-free water (control), 50 ng/mL IFNα (R&D Systems, 10984-IF), 500 ng/mL poly(I:C) (InvivoGen, tlrl-piclv), or CVB3-Woodruff (titer = 1.98 x 10 12 pfu/mL) at a MOI of 20.

Techniques: Marker, Expressing, Derivative Assay, Variant Assay

Journal: bioRxiv

Article Title: Protective IFIH1 variant reduces immune-mediated islet stress and dysfunction in a type 1 diabetes genetic background

doi: 10.64898/2025.12.30.697107

Figure Lengend Snippet: (A-B) , Bar graphs showing the number of differentially expressed genes (DEGs) for each cell type found in (A) MDA5 946T and (B) MDA5 627* SC-islets. (C-E) , Venn diagrams comparing DEGs from both time points for SC-α, -β, and -δ cells from each MDA5 variant treated with either (C) IFNα, (D) poly(I:C), or (E) CVB3. (F-H) , Heatmaps showing log 2 fold change of DEGs unique to (F) SC-α, (G) SC-β, and (H) SC-δ cells from each MDA5 variant following stress treatments.

Article Snippet: Whole SC-islets were then treated with either endotoxin-free water (control), 50 ng/mL IFNα (R&D Systems, 10984-IF), 500 ng/mL poly(I:C) (InvivoGen, tlrl-piclv), or CVB3-Woodruff (titer = 1.98 x 10 12 pfu/mL) at a MOI of 20.

Techniques: Variant Assay

Journal: bioRxiv

Article Title: Protective IFIH1 variant reduces immune-mediated islet stress and dysfunction in a type 1 diabetes genetic background

doi: 10.64898/2025.12.30.697107

Figure Lengend Snippet: (A-B) , Heatmaps depicting log 2 fold change of (A) NFκB signaling-associated and (B) type 1 IFN-associated DEGs across SC-α, -β, and -δ cells when comparing the corresponding IFNα treatment to control or stress-treated MDA5 variants to each other. Positive fold change values (red) in variant-to-variant comparisons correspond to upregulation in MDA5 946T cells when compared to their MDA5 627* counterparts. (C-E) , rt-qPCR of type 1 IFN-associated genes (n = 3). Error bars represent s.e.m.** = Brown-Forsythe and Welch ANOVA tests followed by Dunnett’s T3 multiple comparisons test. (F) , Heatmap depicting log 2 fold chance of apoptosis-associated DEGs across SC-α, -β, and -δ cells when comparing the corresponding stress treatment to control or stress-treated MDA5 variants to each other. Positive fold change values (red) in variant-to-variant comparisons correspond to upregulation in MDA5 946T cells when compared to their MDA5 627* counterparts. (G) , Apoptosis assay measuring caspase 3/7 activity normalized to cell viability (n = 3). Error bars represent s.e.m. ** = Ordinary two-way ANOVA followed by Sidak’s multiple comparisons test. (H) , rt-qPCR of viral genome expression in CVB3-infected SC-islets (n = 3). Error bars represent s.e.m. ** = Unpaired t test. (I) , Intracellular viral titer in SC-islets following 48 h of CVB3 infection (n = 3). Error bars represent s.e.m. ** = Welch’s unpaired t test. Outlier identified in infected MDA5 627* SC-islet samples using the Grubbs’ method.

Article Snippet: Whole SC-islets were then treated with either endotoxin-free water (control), 50 ng/mL IFNα (R&D Systems, 10984-IF), 500 ng/mL poly(I:C) (InvivoGen, tlrl-piclv), or CVB3-Woodruff (titer = 1.98 x 10 12 pfu/mL) at a MOI of 20.

Techniques: Control, Variant Assay, Quantitative RT-PCR, Apoptosis Assay, Activity Assay, Expressing, Infection