Journal: Cell Communication and Signaling : CCS

Article Title: PSPC1-SMAD3 axis regulates iron-induced beiging of adipocytes in white adipose tissue

doi: 10.1186/s12964-025-02570-9

Figure Lengend Snippet: The thermogenic program of beige adipocytes is functionally associated with iron metabolism. A Schematic diagram illustrating the protocol for in vitro beige adipocyte differentiation, ferric ammonium citrate (FAC) treatment (50µM) and RNA-sequencing. B Representative images of Oil Red O staining for lipid droplets in CON and FAC-treated beige adipocytes. Scale bar:50 µm. C Heatmap showing relative mRNA expression of genes related to thermogenesis, mitochondrial function, and fatty acid metabolism in differentiated beige adipocytes treated with vehicle CON or FAC. D Relative mRNA levels of genes related to thermogenesis, fatty acid oxidation, and adipogenesis in CON and FAC-treated beige adipocytes. E Gene Ontology (GO) biological process enrichment analysis of differentially expressed genes identified by RNA-seq between CON and FAC-treated cells. F Immunoblots analysis of UCP1, FABP4, and FTH1 protein levels in CON and FAC-treated beige adipocytes. G Non-heme iron content in different adipose tissues (BAT, iWAT, eWAT) from RT and cold-exposed mice (4°C for 3 days). H Relative mRNA levels of iron metabolism-related genes in iWAT from RT and cold-exposed mice. I Non-heme iron content in different adipose tissues (BAT, iWAT, eWAT) of mice fed a chow diet (CD) or a high-fat diet (HFD) for 12 weeks. J Relative mRNA levels of iron metabolism-related genes in iWAT from CD and HFD-fed mice. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s., not significant

Article Snippet: The membranes were blocked with 5% milk powder in Tris-buffered saline with 0.1% Tween 20 (TBST) at room temperature for 1 h, followed by incubation overnight at 4 °C with the following primary antibody: anti-UCP1 (Abcam, ab234430), anti FTH1 (Abclone, A19544), anti phospho-SMAD3 (Ser423/425) (CST, #9520), anti SMAD3(CST, #9523), anti phospho-SMAD2 (Ser465/467), anti SMAD2 (ZENBIO, 382472), anti FABP4 (Proteintech, 12802-1-AP), anti PSPC1 (Proteintech, 16714-1-AP), anti HSP90(Proteintech, 60318-1-Ig), antiβ-Actin (Abclone, AC026), anti PGC1α (Proteintech, 66369-1-Ig).

Techniques: In Vitro, RNA Sequencing, Staining, Expressing, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: PSPC1-SMAD3 axis regulates iron-induced beiging of adipocytes in white adipose tissue

doi: 10.1186/s12964-025-02570-9

Figure Lengend Snippet: Iron promotes beiging of adipocytes in iWAT and prevents diet-induced obesity in mice. A Schematic diagram of the experimental design. C57BL/6J mice were fed a HFD containing either control levels of iron (CTRL, 37 mg/kg) or enriched with iron (IED, 200 mg/kg) for 10 weeks. B Body weight progression over 10 weeks. C Representative H&E staining of iWAT, eWAT, and BAT sections. Scale bar, 50 μm. D - E Glucose tolerance test (GTT) and insulin tolerance test (ITT) between CTRL and IED group. F Oxygen consumption (VO2) during light and dark cycles in CTRL and IED group. G Relative mRNA levels of thermogenic genes ( Ucp1 , Cidea , Pgc1a ) and iron storage gene ( Fth1 , Ftl1 ) in iWAT. H Representative Western blot of UCP1 and FTH1 protein levels in iWAT. I Schematic diagram of the experimental design. HFD-fed mice received weekly in situ injections of iron dextran (Iron) or vehicle into both iWAT depots for 10 weeks. J Body weight progression over the 10-week treatment period. K H&E staining of BAT, iWAT, and eWAT sections. Scale bar, 50 μm. L - M GTT and ITT curves in vehicle and iron-injected mice. N Representative western blot of UCP1 and FTH1 protein levels in iWAT from vehicle and iron-injected mice. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s., not significant

Article Snippet: The membranes were blocked with 5% milk powder in Tris-buffered saline with 0.1% Tween 20 (TBST) at room temperature for 1 h, followed by incubation overnight at 4 °C with the following primary antibody: anti-UCP1 (Abcam, ab234430), anti FTH1 (Abclone, A19544), anti phospho-SMAD3 (Ser423/425) (CST, #9520), anti SMAD3(CST, #9523), anti phospho-SMAD2 (Ser465/467), anti SMAD2 (ZENBIO, 382472), anti FABP4 (Proteintech, 12802-1-AP), anti PSPC1 (Proteintech, 16714-1-AP), anti HSP90(Proteintech, 60318-1-Ig), antiβ-Actin (Abclone, AC026), anti PGC1α (Proteintech, 66369-1-Ig).

Techniques: Control, Staining, Western Blot, In Situ, Injection

Journal: Cell Communication and Signaling : CCS

Article Title: PSPC1-SMAD3 axis regulates iron-induced beiging of adipocytes in white adipose tissue

doi: 10.1186/s12964-025-02570-9

Figure Lengend Snippet: Iron inhibits the activation of SMAD3 in beige adipocytes. A Schematic diagram illustrating the protocol for ATAC-sequencing in in vitro differentiated beige adipocytes treated with FAC. B ATAC-seq signal distribution near the transcription start sites (TSS). C Pie chart showing the genomic distribution of all identified ATAC-seq peaks. D Motif enrichment analysis of differential ATAC-seq peaks (comparing FAC vs. CON). E Transcription factor footprint (TF) analysis for SMAD2::SMAD3 within differential ATAC-seq peaks. F Relative mRNA levels of Smad2 and Smad3 in CON and FAC-treated beige adipocytes. G , H Protein levels of phosphorylated and total SMAD2 and SMAD3 levels in CON and FAC-treated beige adipocytes. I , J Immunoblots phosphorylated SMAD3 (p-SMAD3), total SMAD3, UCP1, FABP4, and FTH1 in beige adipocytes treated with vehicle (CON), iron chelator deferoxamine (DFO), or DFO + FAC. Data in are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s., not significant

Article Snippet: The membranes were blocked with 5% milk powder in Tris-buffered saline with 0.1% Tween 20 (TBST) at room temperature for 1 h, followed by incubation overnight at 4 °C with the following primary antibody: anti-UCP1 (Abcam, ab234430), anti FTH1 (Abclone, A19544), anti phospho-SMAD3 (Ser423/425) (CST, #9520), anti SMAD3(CST, #9523), anti phospho-SMAD2 (Ser465/467), anti SMAD2 (ZENBIO, 382472), anti FABP4 (Proteintech, 12802-1-AP), anti PSPC1 (Proteintech, 16714-1-AP), anti HSP90(Proteintech, 60318-1-Ig), antiβ-Actin (Abclone, AC026), anti PGC1α (Proteintech, 66369-1-Ig).

Techniques: Activation Assay, Sequencing, In Vitro, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: PSPC1-SMAD3 axis regulates iron-induced beiging of adipocytes in white adipose tissue

doi: 10.1186/s12964-025-02570-9

Figure Lengend Snippet: Inhibition of SMAD3 signaling promotes thermogenesis in beige adipocytes. A Representative Oil Red O staining of beige adipocytes treated with CON, FAC, SMAD3 inhibitor SB-431,542 (2µM) or SB-431,542 plus FAC. B Relative mRNA levels of thermogenic genes in cells treated as in ( A ). C - D Immunoblots of p-SMAD3, SMAD3, UCP1, FABP4, and FTH1 in cells treated as in ( A ). E Representative Oil Red O staining of beige adipocytes treated with CON or SMAD3 inhibitor SIS3. F Relative mRNA levels of thermogenic genes in cells treated as in ( E ). G , H Immunoblots of p-SMAD3, SMAD3, and UCP1 in cells treated as in ( E ). I , J Immunoblots of p-SMAD3/SMAD3 in iWAT from mice in CTRL and IED group (from Fig. ). K , L Immunoblots of p-SMAD3/SMAD3 in iWAT from vehicle and iron-injected mice (from Fig. ). Data in are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s., not significant

Article Snippet: The membranes were blocked with 5% milk powder in Tris-buffered saline with 0.1% Tween 20 (TBST) at room temperature for 1 h, followed by incubation overnight at 4 °C with the following primary antibody: anti-UCP1 (Abcam, ab234430), anti FTH1 (Abclone, A19544), anti phospho-SMAD3 (Ser423/425) (CST, #9520), anti SMAD3(CST, #9523), anti phospho-SMAD2 (Ser465/467), anti SMAD2 (ZENBIO, 382472), anti FABP4 (Proteintech, 12802-1-AP), anti PSPC1 (Proteintech, 16714-1-AP), anti HSP90(Proteintech, 60318-1-Ig), antiβ-Actin (Abclone, AC026), anti PGC1α (Proteintech, 66369-1-Ig).

Techniques: Inhibition, Staining, Western Blot, Injection

Journal: Cell Communication and Signaling : CCS

Article Title: PSPC1-SMAD3 axis regulates iron-induced beiging of adipocytes in white adipose tissue

doi: 10.1186/s12964-025-02570-9

Figure Lengend Snippet: PSPC1 interacts with SMAD3 to regulate its activation and is involved in the regulation of iron on beiging of adipocytes. A Venn diagram illustrating the strategy to identify potential upstream regulators linking iron and SMAD3, integrating RNA-seq, ATAC-seq, published datasets ( GSE35011 , GSE22424 , GSE129573 ), and literature mining, leading to PSPC1. B ATAC-seq bedgraph panels of the Pspc1 locus showing the peak locations relative to the TSS. C Relative mRNA levels of Pspc1 in CON and FAC-treated beige adipocytes, measured by qPCR. D , E Immunoblots of PSPC1 and UCP1 protein levels in CON and FAC-treated cells. F Representative immunofluorescence images showing co-staining for DAPI (blue), SMAD3 (green), and PSPC1 (red) in beige adipocytes. Scale bar, 10 μm. G , H Co-immunoprecipitation (Co-IP) assays showing interaction between endogenous PSPC1 and SMAD3 in beige adipocyte. I , J Representative Oil Red O staining I and mRNA levels of adipogenic/thermogenic genes ( J ) in beige adipocytes transfected with control siRNA (siRNA-ctrl) or Pspc1 siRNA (siRNA-Pspc1). K Immunoblots of PSPC1, p-SMAD3, SMAD3, UCP1, and FABP4 protein levels in cells treated as in ( I ) . L , M Representative Oil Red O staining and relative mRNA levels of adipogenic/thermogenic genes in beige adipocytes transduced with control (CON) or Pspc1 overexpression (PSPC1-OE) lentivirus, with or without subsequent FAC treatment. N Immunoblots of PSPC1, p-SMAD3, SMAD3, UCP1, and FTH1 protein levels in cells treated as in ( L ). Data are presented as mean ± SEM. Scale bar, 50 μm. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s., not significant

Article Snippet: The membranes were blocked with 5% milk powder in Tris-buffered saline with 0.1% Tween 20 (TBST) at room temperature for 1 h, followed by incubation overnight at 4 °C with the following primary antibody: anti-UCP1 (Abcam, ab234430), anti FTH1 (Abclone, A19544), anti phospho-SMAD3 (Ser423/425) (CST, #9520), anti SMAD3(CST, #9523), anti phospho-SMAD2 (Ser465/467), anti SMAD2 (ZENBIO, 382472), anti FABP4 (Proteintech, 12802-1-AP), anti PSPC1 (Proteintech, 16714-1-AP), anti HSP90(Proteintech, 60318-1-Ig), antiβ-Actin (Abclone, AC026), anti PGC1α (Proteintech, 66369-1-Ig).

Techniques: Activation Assay, RNA Sequencing, Western Blot, Immunofluorescence, Staining, Immunoprecipitation, Co-Immunoprecipitation Assay, Transfection, Control, Transduction, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: PSPC1-SMAD3 axis regulates iron-induced beiging of adipocytes in white adipose tissue

doi: 10.1186/s12964-025-02570-9

Figure Lengend Snippet: Iron-induced beiging of adipocytes and metabolic benefits is impaired by PSPC1 overexpression in mice. A Schematic diagram of the in vivo experiment. Mice fed with HFD in control (CTRL) or iron-enriched diet (IED) received iWAT injections of AAV expressing control vector (AAV-Vehicle) or Pspc1 (AAV- Pspc1 ) for a total of 10 weeks HFD feeding. B Body weight in CON and IED mice with AAV-Vehicle/Pspc1 at the end of the study. C Tissue weights of iWAT, eWAT, and BAT expressed as a percentage of total body weight. D Representative H&E staining of BAT, iWAT, and eWAT sections from the four groups. Scale bar, 50 μm. E , F GTT of mice in the four groups. G , H ITT of mice in the four groups. I - J VO2 during light and dark cycles with average values quantified. K Relative mRNA levels of Pspc1 and thermogenic/iron-related genes ( Ucp1 , Cidea , Pgc1a , Fth1 , Ftl1 ) in iWAT from CON and IED group with AAV-Vehicle/Pspc1. L Immunoblots of PSPC1, p-SMAD3, SMAD3, UCP1, and FTH1 protein levels in iWAT. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s., not significant

Article Snippet: The membranes were blocked with 5% milk powder in Tris-buffered saline with 0.1% Tween 20 (TBST) at room temperature for 1 h, followed by incubation overnight at 4 °C with the following primary antibody: anti-UCP1 (Abcam, ab234430), anti FTH1 (Abclone, A19544), anti phospho-SMAD3 (Ser423/425) (CST, #9520), anti SMAD3(CST, #9523), anti phospho-SMAD2 (Ser465/467), anti SMAD2 (ZENBIO, 382472), anti FABP4 (Proteintech, 12802-1-AP), anti PSPC1 (Proteintech, 16714-1-AP), anti HSP90(Proteintech, 60318-1-Ig), antiβ-Actin (Abclone, AC026), anti PGC1α (Proteintech, 66369-1-Ig).

Techniques: Over Expression, In Vivo, Control, Expressing, Plasmid Preparation, Staining, Western Blot

Journal: Journal of Cardiothoracic Surgery

Article Title: NSUN2 inhibits NCOA4 expression to alleviate ferroptosis and inflammation in sepsis-induced myocardial injury in a m 5 C manner

doi: 10.1186/s13019-025-03554-z

Figure Lengend Snippet: NSUN2 suppressed ferroptosis in LPS-induced SIMI. Cellular A , Fe 2 + level, B , GSH content, and C , relative ROS production in each group were determined by commercial kits; D , Western blot was performed to assess the protein levels of NCOA4, FTH1, and GPX4 in each group Fe 2+ , ferrous iron; GSH, glutathione; ROS, reactive oxygen species; NCOA, nuclear receptor coactivator; FTH1, ferritin heavy chain; GPX, glutathione peroxidase

Article Snippet: The primary antibodies used were rabbit antibodies specific for NCOA4 (1:1000; ab314553; Abcam, Cambridge, MA, USA), FTH1 (1:1000; CSB- PA008485 ; Cusabio Engineering Co., LTD, Wuhan, China), GPX4 (1:1000; ab125066; Abcam), and GAPDH (1:1000; ab8245; Abcam).

Techniques: Western Blot

Journal: Journal of Cardiothoracic Surgery

Article Title: NSUN2 inhibits NCOA4 expression to alleviate ferroptosis and inflammation in sepsis-induced myocardial injury in a m 5 C manner

doi: 10.1186/s13019-025-03554-z

Figure Lengend Snippet: NSUN2 inhibited NCOA4 expression in a m 5 C-dependent manner. A , The expression of NCOA4, FTH1, and GPX4 after NSUN2 overexpression in H9c2 cells was detected by RT-qPCR; B , MeRIP-qPCR assay was performed to detect m 5 C levels of NCOA4, FTH1, and GPX4 in vector and NSUN2 groups; C , RIP assay evaluated the interaction of NSUN2 and NCOA4 in H9c2 cells; D , Random Forest method was used to predict possible m 5 C sites of NCOA4; Dual-luciferase gene reporter assay evaluated the binding of NSUN2 and NCOA4 at sites E , 501, F , 1138, and G , 2917; H , RNA stability assay was used to detect the NCOA4 expression when actinomycin D treated at different time points (0, 4, 8, and 12 h) after NSUN2 overexpression NCOA, nuclear receptor coactivator; FTH1, ferritin heavy chain; GPX, glutathione peroxidase; NSUN, NOL1/NOP2/SUN domain; RT-qPCR, reverse transcription-polymerase chain reaction; MeRIP, Methylated RNA immunoprecipitation; RIP, RNA immunoprecipitation; m 5 C, 5-methylcytosine

Article Snippet: The primary antibodies used were rabbit antibodies specific for NCOA4 (1:1000; ab314553; Abcam, Cambridge, MA, USA), FTH1 (1:1000; CSB- PA008485 ; Cusabio Engineering Co., LTD, Wuhan, China), GPX4 (1:1000; ab125066; Abcam), and GAPDH (1:1000; ab8245; Abcam).

Techniques: Expressing, Over Expression, Quantitative RT-PCR, Plasmid Preparation, Luciferase, Reporter Assay, Binding Assay, Stability Assay, Reverse Transcription, Polymerase Chain Reaction, Methylation, RNA Immunoprecipitation

Journal: Journal of Cardiothoracic Surgery

Article Title: NSUN2 inhibits NCOA4 expression to alleviate ferroptosis and inflammation in sepsis-induced myocardial injury in a m 5 C manner

doi: 10.1186/s13019-025-03554-z

Figure Lengend Snippet: Overexpressing NCOA4 downregulated cell viability and upregulated LDH activity, inflammation, and ferroptosis in LPS-induced SIMI. A , NCOA4 mRNA level was detected by RT-qPCR; B , The cell viability of H9c2 cells in each group was analyzed by CCK-8 assay; C , Determination of LDH enzyme activity in each group; D , The contents of TNF-α, IL-6, and IL-8 in each group was evaluated by ELISA; Cellular E , Fe 2 + level, F , GSH content, and G , relative ROS production in each group were determined by commercial kits; H , The protein levels of NCOA4, FTH1, and GPX4 in each group was analyzed by Western blot RT-qPCR, reverse transcription-polymerase chain reaction; CCK-8, cell counting kit-8; LDH, lactate dehydrogenase; ELISA, enzyme-linked immunosorbent assay; TNF-α, tumor necrosis factor-α; IL, interleukin; Fe 2+ , ferrous iron; GSH, glutathione; ROS, reactive oxygen species; NCOA, nuclear receptor coactivator; FTH1, ferritin heavy chain; GPX, glutathione peroxidase

Article Snippet: The primary antibodies used were rabbit antibodies specific for NCOA4 (1:1000; ab314553; Abcam, Cambridge, MA, USA), FTH1 (1:1000; CSB- PA008485 ; Cusabio Engineering Co., LTD, Wuhan, China), GPX4 (1:1000; ab125066; Abcam), and GAPDH (1:1000; ab8245; Abcam).

Techniques: Activity Assay, Quantitative RT-PCR, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Reverse Transcription, Polymerase Chain Reaction, Cell Counting

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Ferritin Heavy Chain in Triple Negative Breast Cancer: A Favorable Prognostic Marker that Relates to a Cluster of Differentiation 8 Positive (CD8+) Effector T-cell Response *

doi: 10.1074/mcp.M113.037176

Figure Lengend Snippet: Prognostic values of FTH1, FTL, and FTH1/FTL ratio in the MS data set of 126 TNBC tumors. A , Expression of FTH1 is significantly higher in good ( n = 82) prognostic TNBC patients, but there is no significant difference in expression of FTL and FTH1/FTL ratio between the two groups of patients; B , Expression of FTH1 significantly correlates to metastasis-free survival in Kaplan-Meier survival analyses (Q1- Q4 = first, second, third and fourth quartile. HR = hazard ratio, CI = confidence interval).

Article Snippet: Tissue sections of 4 μm were incubated for 1 h at room temperature with anti-FTH1 antibody at a dilution of 1:100 against residues near the C terminus of the protein (rabbit-anti-human monoclonal antibody, clone name: EPR3005Y, GeneTex Inc., Irvine, CA, USA).

Techniques: Expressing

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Ferritin Heavy Chain in Triple Negative Breast Cancer: A Favorable Prognostic Marker that Relates to a Cluster of Differentiation 8 Positive (CD8+) Effector T-cell Response *

doi: 10.1074/mcp.M113.037176

Figure Lengend Snippet: GSEA analyses reveal TNBC prognosis and FTH1-associated gene sets. A , Regulation of mesenchymal (M) subtype specific genes in TNBC patients; B , Regulation of immunomodulatory subtype (IM) specific genes in TNBC patients; C , Overlap between TNBC prognosis-associated pathways and FTH1 expression related pathways.

Article Snippet: Tissue sections of 4 μm were incubated for 1 h at room temperature with anti-FTH1 antibody at a dilution of 1:100 against residues near the C terminus of the protein (rabbit-anti-human monoclonal antibody, clone name: EPR3005Y, GeneTex Inc., Irvine, CA, USA).

Techniques: Expressing

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Ferritin Heavy Chain in Triple Negative Breast Cancer: A Favorable Prognostic Marker that Relates to a Cluster of Differentiation 8 Positive (CD8+) Effector T-cell Response *

doi: 10.1074/mcp.M113.037176

Figure Lengend Snippet: Visualization of the network between TNBC prognosis and FTH1 related pathways in EnrichmentMap software. All the enriched pathways were divided into 4 major subnetworks: cytokine signaling, link between innate and adaptive immunity, adaptive immunity, and cell death. Node size represents the gene-set size. Density of node color is related to enrichment of pathways. Thickness of connective lines indicates the degree of overlap between two gene-sets. Top-ranking pathways based on the normalized enrichment score (NES) were listed under each of the subnetworks.

Article Snippet: Tissue sections of 4 μm were incubated for 1 h at room temperature with anti-FTH1 antibody at a dilution of 1:100 against residues near the C terminus of the protein (rabbit-anti-human monoclonal antibody, clone name: EPR3005Y, GeneTex Inc., Irvine, CA, USA).

Techniques: Software

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Ferritin Heavy Chain in Triple Negative Breast Cancer: A Favorable Prognostic Marker that Relates to a Cluster of Differentiation 8 Positive (CD8+) Effector T-cell Response *

doi: 10.1074/mcp.M113.037176

Figure Lengend Snippet: Over-represented network from all the proteins correlated with FTH1 expression. A , A hierarchical cluster of proteins significantly correlating with FTH1 expression forms two distinct clusters, representing to those which show positive (orange bar) and negative (green bar) correlation. The red and green squares highlight two highly correlated subclusters; B , The red subcluster positively correlates with FTH1 expression; C , The green subcluster negatively correlates with FTH1 expression; D , The protein interaction map from the red subcluster is associated with biological process of “cell morphogenesis and adhesion”; E , The protein interaction map from the green subcluster is associated with the biological process of “cell cycle and proteasome.”

Article Snippet: Tissue sections of 4 μm were incubated for 1 h at room temperature with anti-FTH1 antibody at a dilution of 1:100 against residues near the C terminus of the protein (rabbit-anti-human monoclonal antibody, clone name: EPR3005Y, GeneTex Inc., Irvine, CA, USA).

Techniques: Expressing

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Ferritin Heavy Chain in Triple Negative Breast Cancer: A Favorable Prognostic Marker that Relates to a Cluster of Differentiation 8 Positive (CD8+) Effector T-cell Response *

doi: 10.1074/mcp.M113.037176

Figure Lengend Snippet: Subcellular localization of FTH1 and its prognostic significance. A , Cytoplasmic and nuclear staining of FTH1 on TMA and their corresponding cutoff points for Kaplan-Meier analyses; B , Kaplan-Meier analyses show that >75% of tumor cells positive for cFTH1 was associated with favorable prognosis (left panel), whereas >1% positive nFTH1 was related to unfavorable prognosis of TNBC patients (right panel); C , A significant correlation (Spearman correlation) was observed between total FTH1 as measured by LC-MS and cFTH1 staining (left panel), but not nFTH1 staining.

Article Snippet: Tissue sections of 4 μm were incubated for 1 h at room temperature with anti-FTH1 antibody at a dilution of 1:100 against residues near the C terminus of the protein (rabbit-anti-human monoclonal antibody, clone name: EPR3005Y, GeneTex Inc., Irvine, CA, USA).

Techniques: Staining, Liquid Chromatography with Mass Spectroscopy

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Ferritin Heavy Chain in Triple Negative Breast Cancer: A Favorable Prognostic Marker that Relates to a Cluster of Differentiation 8 Positive (CD8+) Effector T-cell Response *

doi: 10.1074/mcp.M113.037176

Figure Lengend Snippet: Univariate and multivariate analyses of cFTH1 and nFTH1 related to metastasis-free survival (n = 147)

Article Snippet: Tissue sections of 4 μm were incubated for 1 h at room temperature with anti-FTH1 antibody at a dilution of 1:100 against residues near the C terminus of the protein (rabbit-anti-human monoclonal antibody, clone name: EPR3005Y, GeneTex Inc., Irvine, CA, USA).

Techniques: Expressing, Staining

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Ferritin Heavy Chain in Triple Negative Breast Cancer: A Favorable Prognostic Marker that Relates to a Cluster of Differentiation 8 Positive (CD8+) Effector T-cell Response *

doi: 10.1074/mcp.M113.037176

Figure Lengend Snippet: Accumulation of CD8+ T-lymphocytes in tumor regions with expression of cFTH1. A , IHC staining of FTH1, CD3, CD4 and CD8 in TNBC whole tissue sections (yellow arrows: CD4+ T-lymphocytes; red arrows: CD8+ T-lymphocytes); B , Box plots summarize the observation in (A). CD4+/CD8+ ratio was significantly increased in TNBC samples with high expression of cFTH1. Data are presented as the median ± interquartile range (Mann-Whitney U test, n = 15 per group); C , A proposed functional model of cFTH1 in the context of the antigen processing and presentation pathway: Ferritin complex captures intracellular Fe 2+ ions (green dots) and converts them into Fe 3+ ions (yellow dots). Accumulation of Fe 3+ iron in tumor cells increases in response to inflammation, and IFN γ (red dots) can enhance the inflammatory response of tumor cells. Together, inflammatory signals can enhance processing of cytosolic antigens through the proteasome, heat shock proteins (HSP), and antigen peptide transporters (TAP), resulting in antigen presentation by major histocompatibility complex class I (MHC I). This in turn attracts CD8+ T-lymphocytes that recognize tumor cells and induce the apoptotic process. On the other hand, cFTH1 can be transported from the cytoplasm to the nucleus. nFTH1 protects from DNA damage caused by reactive oxygen species (ROS) (dashed arrow). Solid lines: observed interactions; Dashed line: hypothetical events; Green arrows: molecular functions of cFTH1; Red arrow: molecular function of nFTH1; Black arrows: pathways driven by FTH1.

Article Snippet: Tissue sections of 4 μm were incubated for 1 h at room temperature with anti-FTH1 antibody at a dilution of 1:100 against residues near the C terminus of the protein (rabbit-anti-human monoclonal antibody, clone name: EPR3005Y, GeneTex Inc., Irvine, CA, USA).

Techniques: Expressing, Immunohistochemistry, MANN-WHITNEY, Functional Assay, Immunopeptidomics

Journal: Clinical and Translational Medicine

Article Title: Integrated multi‐omics profiling landscape of organising pneumonia

doi: 10.1002/ctm2.1782

Figure Lengend Snippet: Reprogramming of lipid metabolism in alveolar macrophages promotes ferroptosis. (A‒D) Violin plot showing the expression levels of FTL (B) and FTH1 (C) in the 13 clusters of healthy control (HC) and organising pneumonia (OP) tissues from the single‐cell RNA sequencing (scRNA‐seq) data. (E) Network of differentially expressed proteins between CON and siCD36 groups (red represents upregulation and green represents downregulation). (F) Gene set enrichment analysis (GSEA) analysis revealed that the changed proteins were mainly enriched in lung fibrosis, mitochondria‐related metabolism, fatty acid metabolism, immune system, cell death and ferroptosis‐related pathways by knocking down CD36. (G) Heatmap showing the differentially expressed proteins related to ferroptosis in alveolar macrophages by knocking down CD36. (H and I) Western blot assay was used to detect the expression of CD36, SLC3A2, GPX4, FTH1, FTL, CES1 and SCD1 in alveolar macrophages. (J) Electron micrograph of alveolar macrophages of OP mouse model (magnification, 30 000×; red arrow, mitochondrion; red rectangle, nucleus). (K) Quantification of perinuclear mitochondrial area, number of mitochondria per square micron, mitochondrial circularity index, cristae number, cristae area, cristae volume and cristae score from (J). (L) The concentration of Fe 2+ , malondialdehyde (MDA) and glutathione (GSH) in alveolar macrophages of OP mouse model. (M and N) Electron micrograph of alveolar macrophages with siCD36 treatment. Quantification of perinuclear mitochondrial area, number of mitochondria per square micron, mitochondrial circularity index, cristae number, cristae area, cristae volume and cristae score from (M). (O) The levels of Fe 2+ , MDA and GSH in alveolar macrophages with siCD36 treatment. Statistical significance is indicated by asterisks; *** p ≤ .001.

Article Snippet: The PVDF membrane was treated with rabbit anti‐SCD1 polyclonal antibody (1:1000, 28678‐1‐AP, Proteintech Group), rabbit anti‐CD36 polyclonal antibody (1:1000, 18836‐1‐AP, Proteintech Group), rabbit anti‐CES1 polyclonal antibody (1:1000, 16912‐1‐AP, Proteintech Group), rabbit anti‐β‐actin monoclonal antibody (1:1000, 66009‐1 Ig, Proteintech Group), rabbit anti‐FTH1 monoclonal antibody (1:1000, R382071, Chengdu Zen Bioscience) or mouse anti‐FTL monoclonal antibody (1:1000, 220988, Chengdu Zen Bioscience), mouse anti‐GPX4 monoclonal antibody (1:1000, 67763‐1‐Ig, Proteintech Group), rabbit anti‐CD98/SLC3A2 polyclonal antibody (1:1000, 15193‐1‐AP, Proteintech Group), rabbit anti‐vimentin polyclonal antibody (1:1000, 10366‐1‐AP, Proteintech Group), rabbit anti‐N‐cadherin polyclonal antibody (1:1000, 22018‐1‐AP, Proteintech Group) and rabbit anti‐E‐cadherin polyclonal antibody (1:1000, 20874‐1‐AP, Proteintech Group) followed by overnight incubation at 4°C in a shaker.

Techniques: Expressing, Control, RNA Sequencing Assay, Western Blot, Concentration Assay