Journal: Redox Biology

Article Title: p53 and fatty acids collaborate to trigger ferroptosis via the FBXO2-FABP5 axis in colorectal cancer

doi: 10.1016/j.redox.2026.104043

Figure Lengend Snippet: FBXO2 is a transcriptional target of p53. Data are represented as mean ± standard deviation (SD), n = 3. ∗ p < 0.05, ∗∗∗ p < 0.001, ns, not significant. See also . (A and B) qPCR analysis of the expression of FBXO2 mRNA in HCT116 p53+/+ (A) and RKO (B) cells treated with or without 5-fluorouracil (5-FU, 40 μM), Etoposide (20 μM), Nutlin-3 (20 μM), or Alrizomadlin (APG-115, 2 μM) for 24 h (C and D) IB analysis of FBXO2 protein expression in HCT116 p53+/+ (C) and RKO (D) cells treated with or without 5-FU, Etoposide, Nutlin-3, or Actinomycin D (Act.D) for 24 h (E and F) qPCR (E) and IB (F) analyses of FBXO2 expression in HCT116 p53−/− cells transfected with the indicated plasmids. (G – J) qPCR (G and I) and IB (H and J) analyses of FBXO2 expression in HCT116 p53+/+ and RKO cells treated with the siRNAs and agents as indicated. (K and L) IB analysis of FBXO2 protein expression in HCT116 p53−/− (K) and HT-29 (with p53-R273H) (L) cells treated with or without the indicated agents. (M) The schematic of the potential p53-responsive element (p53-RE) on the FBXO2 promoter. (N) The luciferase reporter assay verifies the activation of the FBXO2 promoter by p53. (O) ChIP-qPCR analysis confirms the association of p53 with p53-RE on the promoter.

Article Snippet: Nutlin-3, Cisplatin, 5-fluorouracil (5-FU), actinomycin D (Act.D), Etoposide, Alrizomadlin (APG-115), Olaparib, MG132, Liproxstatin-1 (Lipro-1), Ferrostatin-1 (Ferro-1), RSL3, Z-VAD-FMK, Chloroquine (CQ), and Oxaliplatin (Oxa) were purchased from MedChemExpress (Shanghai, China), Erastin and Arachidonic acid (AA) were purchased from Selleck (Shanghai, China).

Techniques: Standard Deviation, Expressing, Transfection, Luciferase, Reporter Assay, Activation Assay, ChIP-qPCR

Journal: Redox Biology

Article Title: p53 and fatty acids collaborate to trigger ferroptosis via the FBXO2-FABP5 axis in colorectal cancer

doi: 10.1016/j.redox.2026.104043

Figure Lengend Snippet: FBXO2 is a transcriptional target of p53. Data are represented as mean ± standard deviation (SD), n = 3. ∗ p < 0.05, ∗∗∗ p < 0.001, ns, not significant. See also . (A and B) qPCR analysis of the expression of FBXO2 mRNA in HCT116 p53+/+ (A) and RKO (B) cells treated with or without 5-fluorouracil (5-FU, 40 μM), Etoposide (20 μM), Nutlin-3 (20 μM), or Alrizomadlin (APG-115, 2 μM) for 24 h (C and D) IB analysis of FBXO2 protein expression in HCT116 p53+/+ (C) and RKO (D) cells treated with or without 5-FU, Etoposide, Nutlin-3, or Actinomycin D (Act.D) for 24 h (E and F) qPCR (E) and IB (F) analyses of FBXO2 expression in HCT116 p53−/− cells transfected with the indicated plasmids. (G – J) qPCR (G and I) and IB (H and J) analyses of FBXO2 expression in HCT116 p53+/+ and RKO cells treated with the siRNAs and agents as indicated. (K and L) IB analysis of FBXO2 protein expression in HCT116 p53−/− (K) and HT-29 (with p53-R273H) (L) cells treated with or without the indicated agents. (M) The schematic of the potential p53-responsive element (p53-RE) on the FBXO2 promoter. (N) The luciferase reporter assay verifies the activation of the FBXO2 promoter by p53. (O) ChIP-qPCR analysis confirms the association of p53 with p53-RE on the promoter.

Article Snippet: Nutlin-3, Cisplatin, 5-fluorouracil (5-FU), actinomycin D (Act.D), Etoposide, Alrizomadlin (APG-115), Olaparib, MG132, Liproxstatin-1 (Lipro-1), Ferrostatin-1 (Ferro-1), RSL3, Z-VAD-FMK, Chloroquine (CQ), and Oxaliplatin (Oxa) were purchased from MedChemExpress (Shanghai, China), Erastin and Arachidonic acid (AA) were purchased from Selleck (Shanghai, China).

Techniques: Standard Deviation, Expressing, Transfection, Luciferase, Reporter Assay, Activation Assay, ChIP-qPCR

Journal: Cell Death & Disease

Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis

doi: 10.1038/s41419-026-08493-8

Figure Lengend Snippet: Primary hepatocytes were transfected with negative control (NC) or PGC-1α CRISPR activation plasmid (PGC-1α CRISPR ACT) and treated with PA, leading to four groups of the Act-NC, Act-NC + PA, Act-PGC-1α and Act-PGC-1α + PA. A Oil Red O, Perls’ Blue staining and Tim23 Immunohistochemistry. B , B ’ Western blot analyses of the relative levels of hepatic PGC-1α, Tim23, Drp1, P-Drp1 Ser616 , ACSL4, GPX4 and FTH1 to GAPDH. C RT-qPCR analyses of the relative levels of PGC-1α, Drp1, ACSL4, GPX4, TFR1 and FTH1 mRNA transcripts. D The GSH levels. E Images of MitoTracker and MitoSOX staining. F Images of C11-BODIPY 581/591 staining. Data are representative images or expressed as the mean ± SD of each group ( n = 3) from at least three independent treatments. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA).

Techniques: Transfection, Negative Control, CRISPR, Activation Assay, Plasmid Preparation, Staining, Immunohistochemistry, Western Blot, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Characterizing the SASP-Dependent Paracrine Spreading of Senescence Between Human Brain Cell Types

doi: 10.64898/2026.02.10.705129

Figure Lengend Snippet: A) Schematic depicting BulkSignalR pipeline which uses known ligand-receptor interactions and affected downstream pathways to analyze their activation based on our bulk RNAseq data from DMSO and BrdU treated human cell lines (created with BioRender). B) Venn diagram showing the number of receptors inferred from BulkSignalR to be activated across each of the five human cell types. Three receptors were identified in common between astrocytes (purple), endothelial cells (pink), and microglia (yellow) which were the cell types shown to be capable of receiving senescence signals and becoming SA β-gal positive: CXCR7, KREMEN2, and GIPR. Only CXCR7 was expressed in the cell types capable of entering secondary senescence (astrocytes, endothelial cells, microglia) ( , S3B). C) TPM expression values of CXCR7 , its ligand CXCL12 , and DPP4 which cleaves and inactivates CXCL12 in DMSO (grey) and BrdU (red) treated cell lines (n=3 replicates). D) Schematic of the four selected SASP inhibitors mechanisms of action: Bindarit is a CCL2 synthesis inhibitor which prevents p65 activation of the CCL2 gene at the promoter region, ISO-1 is a MIF antagonist, ACT-1004-1239 is a CXCR7 antagonist, and Sitagliptin inhibits DPP4 preventing its action of cleaving and inactivating CXCL12 (created with BioRender). Data was analyzed by two-way ANOVA with Tukey’s multiple comparisons test (C). All graphs show mean with error bars depicting standard deviation (ns, p>0.05, ** p<0.01, *** p<0.001).

Article Snippet: Treatment with CXCR7 antagonist ACT-1004-1239 (MedChemExpress, Catalog No. HY-142617) was utilized to prevent CXCR7-associated communication between senescence inducers and receiving cells.

Techniques: Activation Assay, RNA sequencing, Expressing, Standard Deviation

Journal: bioRxiv

Article Title: Characterizing the SASP-Dependent Paracrine Spreading of Senescence Between Human Brain Cell Types

doi: 10.64898/2026.02.10.705129

Figure Lengend Snippet: A) CCK8 viability assay in astrocytes showing normalized percent cell survival following treatment with DMSO control (grey) or DMSO + Bindarit (pink), DMSO + ISO-1 (blue), DMSO + ACT-1004-1239 (orange), or DMSO + Sitagliptin (green) at various concentrations (n=3 replicates). B) CCK8 viability assay in astrocytes showing normalized percent cell survival following treatment with 100 µM BrdU (red) or BrdU + Bindarit (pink), BrdU + ISO-1 (blue), BrdU + ACT-1004-1239 (orange), or BrdU + Sitagliptin (green) at various concentrations (n=3 replicates). C) CCK8 viability assay in microglia showing normalized percent cell survival following treatment with DMSO control (grey) or DMSO + Bindarit (pink), DMSO + ISO-1 (blue), DMSO + ACT-1004-1239 (orange), or DMSO + Sitagliptin (green) at various concentrations (n=3 replicates). D) CCK8 viability assay in microglia showing normalized percent cell survival following treatment with 100 µM BrdU (red) or BrdU + Bindarit (pink), BrdU + ISO-1 (blue), BrdU + ACT-1004-1239 (orange), or BrdU + Sitagliptin (green) at various concentrations (n=3 replicates). Data analyzed by one-way ANOVA with Dunnett’s correction for multiple comparisons (A-D). All graphs show mean with error bars depicting standard deviation (ns, p>0.05, * p<0.05, ** p<0.01, *** p<0.001).

Article Snippet: Treatment with CXCR7 antagonist ACT-1004-1239 (MedChemExpress, Catalog No. HY-142617) was utilized to prevent CXCR7-associated communication between senescence inducers and receiving cells.

Techniques: Viability Assay, Control, Standard Deviation

Journal: bioRxiv

Article Title: Characterizing the SASP-Dependent Paracrine Spreading of Senescence Between Human Brain Cell Types

doi: 10.64898/2026.02.10.705129

Figure Lengend Snippet: A) Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with 100 µM BrdU (red) along with 200 µM Bindarit (pink), 50 µM ISO-1 (blue), 200 µM ACT-1004-1239 (orange), or 2 µM Sitagliptin (green) (n=6 replicates). B) Quantification of percentage of SA β-gal positive microglia following 7-day treatment with 100 µM BrdU (red) along with 200 µM Bindarit (pink), 50 µM ISO-1 (blue), 200 µM ACT-1004-1239 (orange), or 2 µM Sitagliptin (green) (n=6 replicates). C) Timeline showing treatment with DMSO + Bindarit CM or BrdU + Bindarit CM for 7 days. Timeline showing treatment with DMSO or 100 µM BrdU along with SASP inhibitors (ISO-1, ACT-1004-1239, or Sitagliptin) for 7 days. Features of senescence were analyzed 8 days after the initial plating of cells (created with BioRender). D) Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO + Bindarit CM from astrocytes (grey), BrdU + Bindarit CM from astrocytes (red), DMSO CM from astrocytes + SASP inhibitor (grey), or BrdU CM from astrocytes + SASP inhibitor (red) (n=4 replicates). Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO CM from astrocytes + Bindarit (grey) or BrdU CM from astrocytes + Bindarit (red) (n=4 replicates). E) Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO + Bindarit CM from microglia (grey), BrdU + Bindarit CM from microglia (red), DMSO CM from microglia + SASP inhibitor (grey), or BrdU CM from microglia + SASP inhibitor (red) (n=4 replicates). Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO CM from microglia + Bindarit (grey) or BrdU CM from microglia + Bindarit (red) (n=4 replicates). F) Quantification of percentage of SA β-gal positive microglia following 7-day treatment with DMSO + Bindarit CM from microglia (grey), BrdU + Bindarit CM from microglia (red), DMSO CM from microglia + SASP inhibitor (grey), or BrdU CM from microglia + SASP inhibitor (red) (n=4 replicates). Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO CM from microglia + Bindarit (grey) or BrdU CM from microglia + Bindarit (red) (n=4 replicates). G) Quantification of percentage of SA β-gal positive microglia following 7-day treatment with DMSO + Bindarit CM from astrocytes (grey), BrdU + Bindarit CM from astrocytes (red), DMSO CM from astrocytes + SASP inhibitor (grey), or BrdU CM from astrocytes + SASP inhibitor (red) (n=4 replicates). Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO CM from astrocytes + Bindarit (grey) or BrdU CM from astrocytes + Bindarit (red) (n=4 replicates). Data analyzed by unpaired t-test (A-B) and two-way ANOVA with Tukey’s or Šídák’s multiple comparisons test (D-G). All graphs show mean with error bars depicting standard deviation (ns, p>0.05, * p<0.05, ** p<0.01, *** p<0.001).

Article Snippet: Treatment with CXCR7 antagonist ACT-1004-1239 (MedChemExpress, Catalog No. HY-142617) was utilized to prevent CXCR7-associated communication between senescence inducers and receiving cells.

Techniques: Standard Deviation