Journal: iScience

Article Title: Tet2 modulates ER stress responses related to β-cell death and autoimmunity in diabetes

doi: 10.1016/j.isci.2026.114818

Figure Lengend Snippet: Inhibiting TET activity protects human islets from ER stress, cell death and inflammatory responses Gene expression in human islets (n = three to four experiments, each with 4,000–8,000 islet equivalency (IEQ) from each nondiabetic donor after 48 h incubation with brefeldin A (BFA) (A) or thapsigargin (TG) (B) in the presence or absence of TET inhibitor Bobcat339 (Bobcat). The expression of all 12 tested genes, associated with ER stress and cell death were significantly greater in BFA vs. DMSO (∗∗∗∗ p < 0.0001), but significantly reduced in the presence of BFA+Bobcat (ANOVA with Dunnett’s multiple comparison test, ∗∗∗∗ p < 0.0001, ∗ p < 0.05). The expression of 11 of the 12 tested genes were significantly greater in TG vs. DMSO (∗∗∗∗ p < 0.0001), but significantly reduced in the presence of TG + Bobcat (ANOVA with Dunnett’s multiple comparison test,∗∗∗∗ p < 0.0001, ∗∗ p < 0.01). (C) Percent apoptotic and dead cells measured by flow cytometry in human islets cultured for 24 h with BFA with or without Bobcat339 (ANOVA with Tukey’s multiple comparison test,∗∗∗∗ p < 0.0001, ∗∗ p < 0.01). (D) The release of INS DNA with methylation marks indicating β cell origin 30 in human islets cultured for 24 h with BFA with or without Bobcat339 (∗∗∗∗ p < 0.0001 ANOVA with Sidak’s multiple comparison test). (E) Percentage of dead or Indoleamine 2,3-dioxygenase (IDO+) islet cells measured by flow cytometry after 48 h culture with TNFa, IL-1β, IFNy, and IFNa. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 vs. cytokines+DMSO, ANOVA with Tukey’s multiple comparison test data (mean ± 95% CI). The data shown are the individual experiments and represent the mean ± 95 percent confidence intervals.

Article Snippet: DNA was purified from culture supernatants using the Quick-cfDNA Serum & Plasma Kit (Zymo Research) and then bisulfite treated using the EZ DNA Methylation Kit (Zymo Research).

Techniques: Activity Assay, Gene Expression, Incubation, Expressing, Comparison, Flow Cytometry, Cell Culture, DNA Methylation Assay

Journal: Nature Communications

Article Title: TCL1A mediates DNA methylation defects in recurrent hydatidiform mole with NLRP7 pathogenic variants

doi: 10.1038/s41467-026-69744-y

Figure Lengend Snippet: a Imaging of endogenous TCL1A in the human abnormal oocytes/embryos. Blocked, a pre-blocked TCL1A antibody. The total number of oocytes/embryos in 3 independent experiments is indicated. Scale bar, 20 μm. b Relative mean fluorescence intensity of TCL1A and NLRP7 in nucleus (nc) and subcortex (sc) of GV oocytes and blastomeres. The subcortical fluorescence intensity in GV-stage oocytes was set to 1.0. Error bars, SEM. P -values by an unpaired two-tailed Student’s t -test. c Live-cell imaging of GFP-TCL1A in mouse GV oocytes injected with GFP-TCL1A and NLRP7 variant-mCherry mRNAs. The proportion of oocytes exhibiting a similar GFP pattern is indicated. Scale bar, 20 μm. N ≥ 2 replicates for each group. d Co-IP assays detecting the interactions between TCL1A and human DNMT1, DMT3A, DNMT3B3, or DNMT3L. N = 3 independent replicates. e An assay detecting the effect of TCL1A on the DNMT3A activity. The activity of DNMT3A was set to 1.0. Error bars, SEM. N = 3 independent replicates. P -values by an unpaired two-tailed Student’s t -test. f Co-IP assays detecting the effects of overexpressed TCL1A on DNMT3A-DNMT3B3 or DNMT3A-DNMT3L formation. g Generating dual-inducible mESCs and inducing them into fPSCs. S1, Shield1; Dox, doxycycline. See the Methods section for details. h Violin plots displaying the global DNA methylation levels in the four polyclonal fPSCs. N = 5327 bins. The box spans the 25th–75th percentiles, with whiskers indicating 1.5× interquartile range. Central lines, the median. P -values by an unpaired two-tailed Student’s t -test. i Cumulative bar plot of the methylation levels. High, medium, and low methylation levels are defined as >75%, 10–50%, and <10%, respectively. j The box plots showing the DNA methylation levels in genomic Features. The number of each features is indicated. The box spans the 25th–75th percentiles, with whiskers indicating 1.5× interquartile range. Central lines, the median. k A hypothetical model illustrating how pathogenic NLRP7 variants lead to the failure of de novo DNA methylation in oocytes. PDB: hSCMCcore (8X7V); DNMT3A (6PA7). The image was created in BioRender ( https://BioRender.com/lh55y4f ).

Article Snippet: Libraries were prepared using the EpiArt DNA Methylation Library Kit (Vazyme, NE103) according to the manufacturer’s instructions.

Techniques: Imaging, Fluorescence, Two Tailed Test, Live Cell Imaging, Injection, Variant Assay, Co-Immunoprecipitation Assay, Activity Assay, DNA Methylation Assay, Methylation