af6166  (R&D Systems)

Journal: Redox Biology

Article Title: Persulfidation alters gene regulatory programs and promotes endothelial specification

doi: 10.1016/j.redox.2025.103926

Figure Lengend Snippet: Early mesodermal commitment is accompanied by reduced sulfide metabolic flux and altered protein persulfidation that maintains stemness. (A) 34 S cystine derived polysulfides normalized to iPSCs. (B) False color images showing the intensity of the Daz2:Cy5 persulfidation signal iPSCs and in the three germ layers. bar = 10 μm. Fire pseudocoloring was used to distinguish the signal intensity. F-actin: black, NBF–Cl: red. (C) Volcano plot showing persulfidated proteins in iPSCs or cells directed to the mesodermal lineage. (D) OCT4 activity reporter assay in iPSCs treated with solvent (Sol.) or with 5 μmol/L Na 2 S 3 for 24 h. (E) β-catenin stability in mesodermal cells treated with solvent (Sol.) or with 5 μmol/L Na 2 S 3 for 24 h and representative immunoblot at 24 h. (F) Percentage (%) of EOMES-positive live cells in samples treated as in panel E. 3 iPSC lines were used in 2–3 technical replicates. Graphs include the average of all experiments per line. Paired Student's t-test (C, D, F), One-way ANOVA, Tukey's multiple comparison test (A, B), Two-way ANOVA, Tukey's multiple comparison test (E).

Article Snippet: Intracellular staining for mesodermal specification was performed using anti-EOMES antibody (Cell Signaling Technology, Cat. #14-4877-82, clone WD1928, human reactive).

Techniques: Derivative Assay, Activity Assay, Reporter Assay, Solvent, Western Blot, Comparison