Journal: Cell Communication and Signaling : CCS

Article Title: STING-ERO1 signaling exacerbates PARylation-mediated parthanatos in sepsis

doi: 10.1186/s12964-026-02791-6

Figure Lengend Snippet: STING is essential for ROS production, AIF translocation, and cell death. A Immunoblot analysis of STING pathway and γH2AX, PAR levels in WT, STING⁻/⁻, and TBK1⁻/⁻ RAW264.7 cells treated with DMXAA for the indicated times (hours). B Cell viability assessment by Calcein AM (green, live cells) and PI (red, dead cells) staining in WT, STING⁻/⁻, and TBK1⁻/⁻ RAW264.7 cells treated with DMXAA or ADU for 12 h. Scale bar = 25 μm ( C ) Western blot showing AIF levels in cytoplasmic and nuclear fractions of WT and TBK1⁻/⁻ RAW264.7 cells treated with or without DMXAA. D Quantification of AIF protein levels in cytoplasmic and nuclear fractions. E Representative immunofluorescence images showing AIF (magenta) and nuclei (DAPI, blue) in WT and TBK1⁻/⁻ RAW264.7 cells with or without DMXAA treatment. Scale bar = 10 μm. F , G Detection of reactive oxygen species (ROS) using H2DCFDA staining (green) in WT and STING⁻/⁻ RAW264.7 cells treated with DMXAA or ADU. Scale bar = 50 μm ( H ) Quantification of ROS levels in WT, STING⁻/⁻, and TBK1⁻/⁻ RAW264.7 cells treated with DMXAA. I ROS production after co-treatment with DMXAA and PARP1 inhibitor PJ34 at indicated times (hours). Data were shown as the mean ±SD. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001

Article Snippet: DMXAA (HY-10964), ADU-S100 (HY-12885B), PJ34 (HY-13688 A), 3-Aminobenzamide (HY-12022), Olaparib (HY-10162), N-Acetyl-L-cysteine (HY-B0215), Mitoquinone mesylate (HY-100116 A), GSK2795039 (HY-18950), GLX351322 (HY-100111), and EN460 (HY-12837) were purchased from MedChemExpress.

Techniques: Translocation Assay, Western Blot, Staining, Immunofluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: STING-ERO1 signaling exacerbates PARylation-mediated parthanatos in sepsis

doi: 10.1186/s12964-026-02791-6

Figure Lengend Snippet: The effect of the PARP1 inhibitor PJ34 on inflammation and cell death caused by STING activation. A , B Western blot analysis of STING pathway and PAR level in RAW264.7 (A) or iBMDM (B) cells treated with DMXAA alone or in combination with PARP1 inhibitor (PJ34) for the indicated times (hours). C Evaluate the activation of the IFN luciferase reporter gene in RAW264.7 cells mediated by ADU, with or without PARP1 inhibitors (PJ34, 3AB, Olaparib) or NAC. D Relative mRNA expression levels of cytokines (IFN-β, IL-6, TNF-α, CCL2, ISG15, IL-10, IL-1β, and NOS2) in RAW264.7 cells treated with DMXAA alone or in combination with PJ34. E Immunofluorescence staining of RAW264.7 cells treated with DMXAA for 12 hours, with or without PJ34, using Calcein AM (green, live cells) and PI (red, dead cells). Scale bar = 25 μm. F Quantification of cytotoxicity and cell viability after DMXAA treatment with or without PJ34 for the indicated times. Cytotoxicity was assessed by LDH release, and cell viability by CCK-8 assay. G Subcellular fractionation and immunoblotting showing AIF localization in cytoplasmic and nuclear fractions after DMXAA treatment with or without PJ34. H Quantification of AIF protein levels in cytoplasmic and nuclear fractions. I immunofluorescence images of AIF (green) and nuclei (DAPI, blue) in RAW264.7 cells after DMXAA treatment with or without PJ34. Scale bar = 5 μm. Data were shown as the mean ±SD. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001

Article Snippet: DMXAA (HY-10964), ADU-S100 (HY-12885B), PJ34 (HY-13688 A), 3-Aminobenzamide (HY-12022), Olaparib (HY-10162), N-Acetyl-L-cysteine (HY-B0215), Mitoquinone mesylate (HY-100116 A), GSK2795039 (HY-18950), GLX351322 (HY-100111), and EN460 (HY-12837) were purchased from MedChemExpress.

Techniques: Activation Assay, Western Blot, Luciferase, Expressing, Immunofluorescence, Staining, CCK-8 Assay, Fractionation

Journal: Cell Communication and Signaling : CCS

Article Title: STING-ERO1 signaling exacerbates PARylation-mediated parthanatos in sepsis

doi: 10.1186/s12964-026-02791-6

Figure Lengend Snippet: Inhibition of PARP1 ameliorates tissue damage and mortality caused by sepsis. A Representative HE staining images of intestine and lung tissue after CLP with or without PJ34. Scale bar = 50 μm. B TUNEL staining indicated the presence of dead cells (green fluorescence) in the intestinal tissue of each group. Scale bar = 50 μm. C Quantitative assessment of intestinal histopathological damage scores (chiu’s score) in each group. D Relative mRNA expression levels of IL6 in intestinal tissues of each group. E Survival curves at 14 days after CLP in mice with or without PJ34 treatment. F Western blot analysis of PARP1, PAR, AIF, and MIF protein expression levels in intestinal tissues of each group. G Relative quantification of PARP1, PAR, AIF, and MIF protein expression in intestinal tissues of each group. H The schematic diagram illustrates the molecular mechanism of interaction between STING and ERO1, which leads to parthanatos in sepsis. Data were shown as the mean ±SD. *p<0.05, **p<0.01, ***p<0.001

Article Snippet: DMXAA (HY-10964), ADU-S100 (HY-12885B), PJ34 (HY-13688 A), 3-Aminobenzamide (HY-12022), Olaparib (HY-10162), N-Acetyl-L-cysteine (HY-B0215), Mitoquinone mesylate (HY-100116 A), GSK2795039 (HY-18950), GLX351322 (HY-100111), and EN460 (HY-12837) were purchased from MedChemExpress.

Techniques: Inhibition, Staining, TUNEL Assay, Fluorescence, Expressing, Western Blot, Quantitative Proteomics

Journal: The Journal of Cell Biology

Article Title: MEIS homeodomain proteins facilitate PARP1/ARTD1-mediated eviction of histone H1

doi: 10.1083/jcb.201701154

Figure Lengend Snippet: PARP activity is required for neuronal differentiation and H1 eviction from the Dcx promoter/enhancer. (A) Schematic outline of the experiments shown in B–F. (B–F) Reduced neurogenesis and enhanced astrogliogenesis upon pharmacological PARP inhibition: (B and C) Proportions of neurons (B) and astrocytes (C) generated in the presence of increasing concentrations of Olaparib. (D) Representative images of cultures differentiated in the presence of Olaparib or 0.01% DMSO as control. Arrowheads indicate DCX-positive neuronal processes. (E and F) Neurons and astrocytes generated in the presence of 3AB (E) or PJ34 (F). (G, left) Outline of the experiment; (right) neuronal differentiation after shRNA-mediated knockdown of PARP1. (H, left) Outline of the experiment; (right) qPCR for Meis2 , Pbx1 , and Dcx transcripts in cells differentiated for 10 h by growth factor removal and plating on laminin in the presence of 100 nM Olaparib. Expression is shown relative to expression determined in cells treated with 0.01% DMSO (vehicle only). (I, left) Outline of the experiment; (right) ChIP-qPCR for H1 at Dcx(−2.7) in cells differentiated for 5 h in the presence of 6 mM 3AB (gray bars) or water as vehicle control (black bars). Statistical significance of ChIP results between experimental groups is given as p = numerical value. Data are represented as means ± SEM. Samples sizes and the number of biological replicates are listed in Table S4.

Article Snippet: To inhibit PARP1 enzymatic activity during the differentiation process, the medium was supplemented with PJ34 hydrochloride (Tocris Bioscience), 3AB (Sigma-Aldrich), or Olaparib (AZD2281; Lynparza; Selleckchem) in the listed concentrations.

Techniques: Activity Assay, Inhibition, Generated, Control, shRNA, Knockdown, Expressing, ChIP-qPCR

Journal: The Journal of Cell Biology

Article Title: MEIS homeodomain proteins facilitate PARP1/ARTD1-mediated eviction of histone H1

doi: 10.1083/jcb.201701154

Figure Lengend Snippet: Neuronal differentiation is not compromised when pharmacological inhibition of PARP follows the induction of cellular differentiation. (A, left) Outline of the experiment: differentiation was induced in primary SVZ progenitor cells by growth factor removal and plating on laminin 12 h before addition of PJ34 to the culture medium. Addition of water (diluent) served as control; (right) proportion of TuJ1 + neurons generated under both conditions; after 3 d of differentiation, 53.9 ± 3.6% (SD) of the cells differentiated into DCX + neurons under standard conditions, and 49.9 ± 7.6% (SD) differentiated when PJ34 was added to the medium after the first 12 h of differentiation. (B) Representative micrographs of these experiments. Data are represented as means ± SEM, and the number of biological replicates is listed in Table S4.

Article Snippet: To inhibit PARP1 enzymatic activity during the differentiation process, the medium was supplemented with PJ34 hydrochloride (Tocris Bioscience), 3AB (Sigma-Aldrich), or Olaparib (AZD2281; Lynparza; Selleckchem) in the listed concentrations.

Techniques: Inhibition, Cell Differentiation, Control, Generated