Journal: Redox Biology

Article Title: FOS and JUN regulate oxidative stress and steroidogenesis in human aldosterone-producing adenomas

doi: 10.1016/j.redox.2025.103982

Figure Lengend Snippet: Identification of oxidative stress-responsive genes FOS and JUN in human adrenocortical cells A . Gene set variation analysis (GSVA) of hallmark pathways in APAs and adjacent tissues based on integrated datasets ( GSE60042 , GSE64957 , and GSE156931 ). B . Oxidative stress-related pathways identified by gene set enrichment analysis (GSEA) of APAs versus adjacent tissues, based on integrated datasets ( GSE60042 , GSE64957 , and GSE156931 ). C and D . Volcano plots of RNA sequencing data showing differentially expressed genes (DEGs) in HAC15 adrenocortical cells treated with 4 μM RSL-3 (oxidative stressor via GPX4 inhibition and lipid peroxidation) or 250 μM H 2 O 2 (direct oxidant) for 3h vs. DMSO-treated controls (n = 3 biological replicates). Dashed lines: adjusted p < 0.05; |log2 fold change| > 1. E . Venn diagram identifying overlapping DEGs between RSL-3/H 2 O 2 -treated HAC15 cells and spatial transcriptomics (ST) data from aldosterone-producing adenomas (APAs, n = 7) vs. paired adjacent adrenal cortex. F . Gene Ontology (GO) enrichment of overlapping genes in Panel B, highlighting oxidative stress and steroidogenic pathways. NES, normalized enrichment score; FDR, false discovery rate; NS, not significant.

Article Snippet: Real-time PCR amplification was performed on the BIO-RAD, CFX Duet PCR system using the following TaqMan probes from Thermo Fisher Scientific: FOS (Hs00170630_m1), JUN (Hs99999141_s1), GAPDH (Hs02786624_g1), CYP11B2 (Hs01597732_m1), CYP11B1 (Hs01596406_gH) or StAR (Hs00986559_g1).

Techniques: RNA Sequencing, Inhibition

Journal: Redox Biology

Article Title: FOS and JUN regulate oxidative stress and steroidogenesis in human aldosterone-producing adenomas

doi: 10.1016/j.redox.2025.103982

Figure Lengend Snippet: Validation of FOS and JUN expression and oxidative stress activity in APAs and paired adjacent adrenal tissues A. Boxplots showing the expression levels of CYP11B2 , FOS , and JUN across APAs and paired adjacent adrenal tissues, integrating data from GSE60042 , GSE64957 , and GSE156931 . B . Spatial transcriptomics heatmaps of FOS, JUN, CYP11B2, and KCNJ5 across APA-adrenal cortex pairs (n = 7). C . Representative Spatial transcriptomics expression maps of CYP11B2 , FOS , and JUN in KCNJ5 -mutant ( KCNJ5 MUT ) vs. wild-type APAs ( KCNJ5 WT ) and adjacent tissues. D . Integrated UMAP embedding and boxplot showing the distribution of oxidative stress scores in seven APAs and their paired adjacent tissues. E . Representative spatial feature plots of oxidative stress scores projected onto H&E-stained APA and paired adjacent tissue sections. F and G . Gene expression analysis of FOS and JUN in APAs and paired adjacent adrenal tissues from 10 KCNJ5 -mutant and 10 wild-type APA patients. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Real-time PCR amplification was performed on the BIO-RAD, CFX Duet PCR system using the following TaqMan probes from Thermo Fisher Scientific: FOS (Hs00170630_m1), JUN (Hs99999141_s1), GAPDH (Hs02786624_g1), CYP11B2 (Hs01597732_m1), CYP11B1 (Hs01596406_gH) or StAR (Hs00986559_g1).

Techniques: Biomarker Discovery, Expressing, Activity Assay, Mutagenesis, Staining, Gene Expression

Journal: Redox Biology

Article Title: FOS and JUN regulate oxidative stress and steroidogenesis in human aldosterone-producing adenomas

doi: 10.1016/j.redox.2025.103982

Figure Lengend Snippet: Differential expression and activation of FOS and JUN in adrenal tumors and adjacent adrenal cortex A . Immunohistochemistry of adrenal sections from APAs with mutations (MUT) in KCNJ5 , CACNA1D , ATP1A1 or ATP2B3 . Sections were stained with hematoxylin and eosin (H&E) and immunostained for CYP11B2 (aldosterone synthase), total FOS, phospho-FOS (Ser 32; activated form), total JUN, and phospho-JUN (Ser 73; activated form). B . Immunohistochemistry of adrenal sections from cortisol-producing adenomas (CPAs), non-functional adrenal incidentalomas, and histologically normal adrenal cortex (from nephrectomy specimens). Sections were stained with hematoxylin and eosin (H&E) and immunostained for total FOS, phospho-FOS (Ser 32; activated form), total JUN, and phospho-JUN (Ser 73; activated form). Three biologically independent specimens were analyzed per group. zG: zona glomerulosa; zF: zona fasciculata.

Article Snippet: Real-time PCR amplification was performed on the BIO-RAD, CFX Duet PCR system using the following TaqMan probes from Thermo Fisher Scientific: FOS (Hs00170630_m1), JUN (Hs99999141_s1), GAPDH (Hs02786624_g1), CYP11B2 (Hs01597732_m1), CYP11B1 (Hs01596406_gH) or StAR (Hs00986559_g1).

Techniques: Quantitative Proteomics, Activation Assay, Immunohistochemistry, Staining, Functional Assay

Journal: Redox Biology

Article Title: FOS and JUN regulate oxidative stress and steroidogenesis in human aldosterone-producing adenomas

doi: 10.1016/j.redox.2025.103982

Figure Lengend Snippet: Oxidative stress suppresses steroidogenesis and induces FOS and JUN expression and phosphorylation in human adrenocortical cells A and B . Intracellular ROS level and cell viability in HAC15 cells treated with DMSO, 4 μM RSL-3 or 10 μM Lip-1 + 4 μM RSL-3 for 4h (n = 6 independent experiments). C and D . ROS level and cell viability in HAC15 cells treated with DMSO or 250 μM H2O2 for 4h (n = 6). E. Aldosterone secretion in HAC15 cells treated with DMSO, 10 nM Ang II, 10 nM Ang II + 4 μM RSL-3, or 10 nM Ang II + 250 μM H 2 O 2 (n = 3). F. Cortisol production in HAC15 cells treated with DMSO, 10 μM FSK, 10 μM FSK +4 μM RSL-3, or 10 μM FSK +250 μM H 2 O 2 (n = 3). G . Gene expression analysis of FOS and JUN in HAC15 cells treated with DMSO, 4 μM RSL-3, 10 μM Lip-1 + 4 μM RSL-3 or 250 μM H2O2 for 4h (n = 6). H . Cyto-immunofluorescence analysis of total FOS, phospho-FOS (Ser 32), total JUN, and phospho-JUN (Ser 73) in expression in HAC15 cells treated with DMSO, 4 μM RSL-3, 10 μM Lip-1 + 4 μM RSL-3 or 250 μM H2O2 for 4h (n = 3). Nuclei were counterstained with DAPI. Data in panels A–H were normalized to the DMSO control group. Statistical significance was determined by Welch's ANOVA with Games-Howell's post-test (panels A and B), Student's t -test (panels C and D), or one-way ANOVA with Dunnett's post-test (panels E–H). Data are shown as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant. FSK: Forskolin.

Article Snippet: Real-time PCR amplification was performed on the BIO-RAD, CFX Duet PCR system using the following TaqMan probes from Thermo Fisher Scientific: FOS (Hs00170630_m1), JUN (Hs99999141_s1), GAPDH (Hs02786624_g1), CYP11B2 (Hs01597732_m1), CYP11B1 (Hs01596406_gH) or StAR (Hs00986559_g1).

Techniques: Expressing, Phospho-proteomics, Gene Expression, Immunofluorescence, Control

Journal: Redox Biology

Article Title: FOS and JUN regulate oxidative stress and steroidogenesis in human aldosterone-producing adenomas

doi: 10.1016/j.redox.2025.103982

Figure Lengend Snippet: FOS and JUN modulate the steroidogenic and oxidative stress response in human adrenocortical cells A and B . Time-course analysis of FOS and JUN mRNA expression in HAC15 cells treated with 10 nM Ang II or 10 μM forskolin (n = 3 independent experiments). C and D . Gene (real-time PCR, n = 6) and protein (Western blot analysis, n = 3) expression of FOS and JUN in HAC15 cells after FOS and JUN co-transfection. E . Baseline expression of StAR , CYP11B1 and CYP11B2 in HAC15 cells with FOS and JUN co-overexpression (n = 6). F . CYP11B1 and CYP11B2 gene expression in HAC15 cells with FOS and JUN co-overexpression treated with 10 nM Ang II or 10 μM forskolin for 24 h (n = 6). G . Decrease in cortisol and aldosterone secretion in FOS and JUN co-overexpressing HAC15 cell supernatants treated with DMSO, 10 nM Ang II or 10 μM forskolin for 24 h (n = 6). H . ROS level in FOS and JUN co-overexpressing HAC15 cells treated with or without 4 μM RSL-3 for 4 h (n = 6). Data were normalized to the corresponding control group: A and B vs. DMSO, C-E vs. empty vector, F–H vs. empty vector (EV) + DMSO. Statistical significance was determined by one-way ANOVA with Dunnett's post-test relative to 0 h (panels A and B), Student's t -test (panels C and E), Welch's t -test (panel D) or two-way ANOVA with Sidak's post-test (panels F–H). Data are presented as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant.

Article Snippet: Real-time PCR amplification was performed on the BIO-RAD, CFX Duet PCR system using the following TaqMan probes from Thermo Fisher Scientific: FOS (Hs00170630_m1), JUN (Hs99999141_s1), GAPDH (Hs02786624_g1), CYP11B2 (Hs01597732_m1), CYP11B1 (Hs01596406_gH) or StAR (Hs00986559_g1).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Cotransfection, Over Expression, Gene Expression, Control, Plasmid Preparation

Journal: bioRxiv

Article Title: Human iPSC-derived CNS and retinal microvasculature-on-a-chip models recapitulate hallmarks of diabetic microvascular pathology

doi: 10.64898/2026.01.15.699708

Figure Lengend Snippet: Gene expression changes in CNS-mv-on-a-chip induced by high glucose level. (A) The principal component analysis (PCA) plot shows clustering of samples cultured under physiological (5.5 mM glucose) and hyperglycemic (75 mM glucose) conditions. (B) Volcano plot illustrating differentially expressed genes between physiological and hyperglycemic CNS-mv-on-a-chip conditions. Orange and blue dots represent significantly upregulated and downregulated genes, respectively (FDR < 0.05, |FC| >2). (C) Heatmap showing upregulated genes associated with inflammation. Values are shown as fold changes relative to control group mean. (D) Over-representation analysis (ORA) plot of genes upregulated under hyperglycemia conditions. (E) Quantitative PCR analysis of IL1B, JUN and NFKBIA expression in ECs and PCs isolated from CNS-mv-on-a-chip cultured under physiological 5.5 mM glucose concentration (5.5G), hyperglycemic conditions 25 mM (25G) and 75 mM glucose (75G), or corresponding osmotic controls with mannitol (19.5M and 69.5M). Gene expression was normalized to GAPDH and HMBS as housekeeping genes and calculated using the ΔΔCt method. Data are presented as mean ± SD, n = 3-4.

Article Snippet: First, a pre-amplification step with TaqMan primers (Thermo Fisher Scientific) was carried out for GAPDH (Hs99999905_m1), HMBS (Hs00609297_m1), IL1B (Hs01555410_m1), JUN (Hs01103582_s1), and NFKBIA (Hs00355671_g1).

Techniques: Gene Expression, Cell Culture, Control, Real-time Polymerase Chain Reaction, Expressing, Isolation, Concentration Assay