Journal: Translational Oncology

Article Title: REST-driven upregulation of SFXN3 promotes AML progression via Wnt/β-catenin activation and confers decitabine resistance

doi: 10.1016/j.tranon.2026.102705

Figure Lengend Snippet: REST Regulates SFXN3 Expression by Directly Binding to Its Promoter Region. (A) Identification of transcription factors potentially regulating SFXN3 expression by integrating data from the JASPAR, GTEx, ChIP Atlas, and ENCODE databases. A Venn diagram highlights TFAP2C and REST as common candidates, suggesting them as potential upstream regulators. (B) Quantitative mRNA analysis reveals that REST is significantly upregulated in AML tissues compared to adjacent non-tumor tissues. (C) Spearman correlation analysis shows a strong positive correlation between REST and SFXN3 expression. (D) RT-PCR analysis confirms the knockdown efficiency of two shRNA constructs targeting REST (sh-REST#1 and sh-REST#2), and evaluates their impact on SFXN3 mRNA levels. (E) Western blot analysis validates the protein-level knockdown efficiency of sh-REST#1 and sh-REST#2, along with their effects on SFXN3 protein expression. (F) Quantification of REST and SFXN3 protein expression levels. (G) Prediction of REST binding sites in the SFXN3 promoter using JASPAR and ENCODE databases, with the highest affinity region localized between positions −415 and −395. (H) Binding motif analysis from the JASPAR database confirms a predicted REST binding site within the SFXN3 promoter. A mutant version of this binding site was designed for subsequent experiments. (I) ChIP-seq data demonstrates a prominent REST binding peak at the −415 to −395 region of the SFXN3 promoter. (J) Molecular docking simulation reveals that the DNA-binding domain of REST forms a stable complex with the specific binding site in the SFXN3 promoter. (K) ChIP-qPCR analysis confirms significant enrichment of REST at the SFXN3 promoter, which is markedly reduced following REST knockdown. (L) Dual-luciferase reporter assays show that REST significantly enhances the activity of the wild-type SFXN3 promoter, while mutation of the binding site substantially diminishes this effect.

Article Snippet: Subsequently, luciferase activity was measured using a Dual-Luciferase Reporter Assay Kit (#DL101, Vazyme, Nanjing, China).

Techniques: Expressing, Binding Assay, Reverse Transcription Polymerase Chain Reaction, Knockdown, shRNA, Construct, Western Blot, Mutagenesis, ChIP-sequencing, ChIP-qPCR, Luciferase, Activity Assay

Journal: iScience

Article Title: Exploring roles of neuroglobin in oxygen homeostasis and neurodevelopment based on human cerebral organoid models

doi: 10.1016/j.isci.2026.114997

Figure Lengend Snippet: Experimental workflow and impact of NGB silencing on cerebral organoids development Top: Schematic timeline illustrating the stages of cerebral organoid generation from human-induced pluripotent stem cells (hiPSCs), including embryoid body (EB) formation, neural induction, fusion and expansion, and maturation. All measurement points for data analysis were denoted. Bottom: Experimental workflow comparing control hiPSC-derived cerebral organoids with NGB -silenced organoids. Organoids were analyzed using single-cell RNA sequencing (scRNA-seq) for genotypic profiling, FD-FLIM for functional analysis, confocal microscopy for structural assessment, and microelectrode array (MEA) recordings for functional evaluation.

Article Snippet: The microscope was equipped with a high-power LED excitation source (470 nm nominal wavelength; Thorlabs, M470LP-C2) and a dual-tap CMOS FLIM camera (PCO.FLIM, Excelitas Technologies Corp., Waltham, MA).

Techniques: Control, Derivative Assay, Single Cell, RNA Sequencing, Functional Assay, Confocal Microscopy, Microelectrode Array