Journal: Cell Death & Disease

Article Title: Endoplasmic reticulum stress related super-enhancers suppress cuproptosis via glycolysis reprogramming in lung adenocarcinoma

doi: 10.1038/s41419-025-07613-0

Figure Lengend Snippet: A Copper levels in LUAD tumor tissues ( n = 30) were significantly higher than in matched normal tissues ( n = 30), as determined by a colorimetric copper assay. Absorbance at 580 nm was measured. B Confocal fluorescence imaging revealed increased copper fluorescence (500 nm) in LUAD cell lines (A549, PC9, H2087, and H1975) compared to normal alveolar epithelial cells (BEAS-2B) following a 10 h incubation with 10 µM CuCl 2 . Fluorescence intensity was quantified and normalized to cell area ( n = 3). C Box plot illustrating the relative copper (Cu) concentrations in fresh LUAD tissues, showing significantly higher copper levels in the high-grade group ( n = 15) compared to the low-grade group ( n = 15). D Pearson correlation analysis revealed a positive correlation between Ki-67 expression and copper (Cu) concentration ( n = 30, R = 0.322, p = 0.0271). A simple linear regression line is included to illustrate the relationship. E Representative IHC images of Ki-67 staining in low- and high-grade LUAD tissues. F Box plot illustrates the distribution of H-scores for DLAT and FDX1 expression levels, as determined by immunofluorescence staining on a LUAD tissue microarray ( n = 80). The H-score was calculated as: 1×(percentage of cells with staining score 1) + 2×(percentage of cells with staining score 2) + 3×(percentage of cells with staining score 3). The results show that FDX1 expression is significantly higher in the low-grade group compared to the high-grade group, while no significant difference is observed for DLAT expression. G t-SNE clustering analysis of single-cell RNA sequencing data reveals ten distinct cellular subpopulations. The frequency distribution of different cellular subpopulations in each LUAD sample H and the comparison of subpopulation frequencies between low- and high-grade LUAD groups ( I ). The data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Antibodies used in this study include: XBP1s (Cell Signaling Technology, #40435, 1:1000 for WB, 1:300 for IF, 1:50 for CUT&Tag); XBP1s (Merck, #MABC521, 1:400 for IHC); DLAT (Proteintech, # 68303-1-Ig, 1:400 for IHC and 1:1000 for WB); FDX1 (Proteintech, # 12592-1-AP, 1:400 for IHC); P300 (Cell Signaling Technology, #D2X6N, 1:300 for IF, 1:50 for CUT&Tag); MED1 (Thermo Fisher, #PA5-36114, 1:300 for IF, 1:50 for CUT&Tag); BRD4 (Cell Signaling Technology, #13440, 1:300 for IF, 1:50 for CUT&Tag); RNA Pol II-S2P (Millipore, cat# 04-1571, 1:300 for IF); RNA Pol II-S5P (Millipore, cat# 04-1572, 1:300 for IF); H3K4me3 (Cell Signaling Technology, cat# 9751, 1:300 for IF); H3K27ac (PTMbio, #PTM-160, 1:300 for IF, 1:50 for CUT&Tag); H3K4me1 (PTMbio, #PTM-5158, 1:300 for IF); MGRN1 (Proteintech, #11285-1-AP, 1:2000 for WB, 1:200 for IHC); β-actin (Proteintech, # 81115-1-RR, 1:10000 for WB); LIPT1 (Thermo Fisher, #PA5-106991, 1:1000 for WB and 1:200 for IHC); anti-DYKDDDDK tag (Proteintech, # 80010-1-RR, 1:5000 for WB); anti-HA tag (Proteintech, # 81290-1-RR, 1:5000 for WB); anti-His tag (Proteintech, #66005-1-Ig, 1:5000 for WB); HRP-conjugated goat anti-rabbit (Proteintech, # SA00001-2, 1:10000 for WB); HRP-conjugated goat anti-mouse (Proteintech, # SA00001-1, 1:10000 for WB); CoraLite® Plus 488-Goat Anti-Mouse Recombinant Secondary Antibody (H + L) (Proteintech, # RGAM002, 1:1000 for IF); CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H + L) (Proteintech, # RGAM004, 1:1000 for IF); CoraLite® Plus 647-Goat Anti-Mouse Recombinant Secondary Antibody (H + L) (Proteintech, # RGAM005, 1:1000 for IF)

Techniques: Fluorescence, Imaging, Incubation, Expressing, Concentration Assay, Staining, Immunofluorescence, Microarray, RNA Sequencing, Comparison

Journal: medRxiv

Article Title: Metabolome-wide association of carotid intima media thickness identifies FDX1 as a determinant of cholesterol metabolism and cardiovascular risk in Asian populations

doi: 10.1101/2024.05.14.24307316

Figure Lengend Snippet: 3BH5C, 3beta-hydroxy-5-cholestenoate; eQTL, expression Quantitative Trait Loci; HAoSMCs, Human Aortic Smooth Muscle Cells; H1-hESCs, H1-derived Human Embryonic Stem Cells; HBOs, Hepatobiliary Organoids; FDX1, Ferredoxin-1; KO, Knock-out; MS, Mass Spectroscopy; mQTL, metabolite Quantitative Trait Loci; OE, Over-expression; pQTL, protein Quantitative Trait Loci. Image partially created using Biorender.com

Article Snippet: 24 hours post-seeding, cells underwent transfection with siRNA targeting human FDX1 (Thermo Fisher Scientific, cat no. 4392420, siRNA ID: s223529).

Techniques: Expressing, Derivative Assay, Knock-Out, Mass Spectrometry, Over Expression

Journal: medRxiv

Article Title: Metabolome-wide association of carotid intima media thickness identifies FDX1 as a determinant of cholesterol metabolism and cardiovascular risk in Asian populations

doi: 10.1101/2024.05.14.24307316

Figure Lengend Snippet: (a) Chromosome 11 regional plot showing genetic variants associated with plasma levels of metabolite 3BH5C in HELIOS. (b) Chromosome 11 regional plot showing genetic variants associated with expression of cis -gene FDX1 in whole blood in HELIOS (colocalization posterior probability=0.58). (c) Chromosome 11 regional plot showing genetic variants associated with expression of cis -gene FDX1 in GTEx v8 Whole Blood tissue (colocalization posterior probability= 0.72). (d) Chromosome 11 regional plot showing genetic variants associated with plasma protein abundance of FDX1 in UKBB (colocalization posterior probability= 0.87). In all four regional plots, each dot represents a genetic variant, the genomic positions based on GRCh38 are on the x-axes and -log 10 P-values of associations are on the y-axes. Genes annotated to this region are shown at the bottom of the panel. The top cis -eQTL of FDX1 , rs2846734 is highlighted. The colour of variants indicate their linkage disequilibrium (LD r 2 ) with the top variant in each panel. 1000 Genomes East Asian population served as the LD reference panel for HELIOS panels, 1000 Genomes European population served as LD reference panel for GTEx panel, and the overall 1000 Genomes population served as LD reference panel for UKBB panel which utilized the combined cis -pqTL summary data. Figures were generated using LocusZoom. 3BH5C, 3beta-hydroxy-5-cholestenoate.

Article Snippet: 24 hours post-seeding, cells underwent transfection with siRNA targeting human FDX1 (Thermo Fisher Scientific, cat no. 4392420, siRNA ID: s223529).

Techniques: Clinical Proteomics, Expressing, Quantitative Proteomics, Variant Assay, Generated

Journal: medRxiv

Article Title: Metabolome-wide association of carotid intima media thickness identifies FDX1 as a determinant of cholesterol metabolism and cardiovascular risk in Asian populations

doi: 10.1101/2024.05.14.24307316

Figure Lengend Snippet: (a) FDX1 protein expression in FDX1 knock-out ( FDX1 KO) and FDX1 over-expression ( FDX1 OE) human hepatoma Huh7 cells, FDX1 KO mouse hepatocyte-derived AML12 cells, and FDX1 KO human monocyte-derived THP1 cells, compared to scrambled controls (SC). Metabolite 3BH5C levels in FDX1 KO cells compared to SC in (b) human hepatoma Huh7 cells, (c) mouse hepatocyte-derived AML12 cells, and (d) human monocyte-derived THP1 cells with and without cholesterol treatment. (e) FDX1 protein expression in FDX1 WT and KO, followed by over-expression (OE) and rescue, show robust and stable transgene expression following macrophage differentiation. Measurement of FDX1 levels, and metabolite 3BH5C levels in FDX1 KO compared to wild-type (WT) in (f) H1-derived hepatobiliary organoids (H1-HBOs), and (g) H1-derived macrophages. (h) Measurement of 3BH5C levels upon FDX1 OE and recovery in H1-derived macrophages compared to wild-type (WT). (i) Measurement of 3BH5C levels upon FDX1 OE in Huh 7 cells compared to GFP controls. (j) Quantitative RT-PCR of FDX1 expression in FDX1 knock-down (KD) aortic smooth muscle cells compared to scrambled control. (k) Percentage cholesterol efflux in FDX1 KD cells compared to scrambled control with and without treatment with a positive efflux inducer. *P<0.05, **P<0.01, ***P<0.001 n.s : non significant. 3BH5C, 3beta-hydroxy-5-cholestenoate; FDX1, Ferredoxin-1

Article Snippet: 24 hours post-seeding, cells underwent transfection with siRNA targeting human FDX1 (Thermo Fisher Scientific, cat no. 4392420, siRNA ID: s223529).

Techniques: Expressing, Knock-Out, Over Expression, Derivative Assay, Quantitative RT-PCR, Knockdown, Control