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: Frontiers in Molecular Neuroscience

Article Title: Alteration in Intracellular Zn 2+ Homeostasis as a Result of TRPM2 Channel Activation Contributes to ROS-Induced Hippocampal Neuronal Death

doi: 10.3389/fnmol.2017.00414

Figure Lengend Snippet: TRPM2 channel is crucial for H 2 O 2 -induced production of mitochondrial reactive oxygen species (ROS) in hippocampal neurons. (A,C) Representative fluorescence images showing MitoTracker Red CM-H 2 Xros (MitoROS) fluorescence in WT (left column) or TRPM2-KO (right column) neurons under control (CTL) and treatment with 100 μM or 300 μM H 2 O 2 for 2 h (A) , or in WT neurons treated with 1 μM PJ34 or 1 μM TPEN, 30 min before and during exposure to 300 μM H 2 O 2 (C) . Scale bar is 100 μm. (B,D) Mean MitoROS fluorescence intensity from 3 to 5 independent experiments with 35–70 neurons examined in each experiment, presented relative to the fluorescence level in control neurons in matched experiments. 2-APB was used at 100 μM. * p < 0.05 and *** p < 0.005 indicate significant difference from control. ††† p < 0.005 indicates significant difference from neurons exposed to 300 μM H 2 O 2 alone.

Article Snippet: PJ34 was from Calbiochem, and TPEN from StressMarq Biosciences.

Techniques: Fluorescence

Journal: Nature Communications

Article Title: SPINDOC binds PARP1 to facilitate PARylation

doi: 10.1038/s41467-021-26588-y

Figure Lengend Snippet: a, b Hela cells were cotransfected with GFP-SPINDOC and RFP-PARP1 nanobody plasmids, 24 h later, cells were nontreated ( a ), or treated with PJ34 10 µM an hour ( b ), before being subjected to 355 nm UVA laser microirradiation-coupled live-cell imaging. Cells were treated with 200 ng/ml Hoechst 30 mins for presensitization prior to laser damage and imaging. Confocal images (40X) were taken every 5 s for 10 mins and representative images of the irradiation-path signals are shown. c Quantification of the corecruitment of GFP-SPINDOC and RFP-αPARP1 in a and b . Graphs represent mean ± SD, n = 10 biologically independent cells. d Hela-WT cells, Hela-SPINDOC KO cells, and the same cells rescued with GFP-SPINDOC and GFP-SPINDOC∆115–165aa mutant were transfected with RFP-αPARP1 nanobody plasmid and incubated for 24 h, before presensitizing with Hoechst (200 ng/ml), 355-nm UVA laser-induced damage and confocal imaging (40X). Images were taken every 20 s for 15 mins. e Quantification of RFP- αPARP1 recruitment in d . All timepoints are normalized to the peak of maximal recruitment of RFP-PARP1 to a value of 1.0 by each group. Graphs represent mean ± SD, n = 10 biologically independent cells. One-tailed Student t- tests were performed at 400 s, comparing Hela-WT with KO, P = 0.0118; Hela-KO vs KO-SPINDOC rescue, P = 0.0480; and Hela-KO vs KO - SPINDOC mutant, P = 0.0838. f Hela-SPINDOC WT and KO cell lines were treated with 10 Gy IR, allowed to recover for 0.5, 3, and 6 h, and subjected to Western blot analysis using the indicated antibodies. The data are representative of three independent experiments. g An in vitro PARylation assay was performed in the presence of purified GST, GST-SPINDOC or GST-SPINDOC115–165 proteins, NAD + , and recombinant PARP1, at room temperature for 30 min. Auto-PARylation signal was detected with αPAR antibody. The data are representative of three independent experiments. h PARP1 transcription level was evaluated by RT-qPCR in SPINDOC KO and overexpressed cells. One-tailed Student t -tests were performed, SPINDOC KO vs WT, P = 0.1822; Flag-SPINDOC vs vector, P = 0.2270. i Hela-SPINDOC WT and KO cell lines were treated with 10-Gy IR, allowed to recover for 0.5, 3, and 6 h, and subjected to RT-qPCR assay for the six indicated targets. Graphs represent mean ± SD, n = 3 biologically independent samples. One-tailed Student t -tests were performed, SPINDOC WT, ** P = 0.0044, * P = 0.0121, *** P < 0.001, **** P < 0.0001. SPINDOC KO, RAD51: * P = 0.0382, ** P = 0.0091. NRF1: * P = 0.1027, 0.0871, 0.0302. NF-κB1: P = 0.0018, 0.0093, 0.3797; P 130: P = 0.0304, 0.0054, 0.0066; hNIS: P = 0.0154, 0.0068, 0.0030. j Hela-SPINDOC KO cells rescued with GFP-SPINDOC, pEGFP-C1 as a control, were treated with 10-Gy IR, allowed to recover for 0.5, 3, and 6 h, and subjected to RT-qPCR assay. Graphs represent mean ± SD, n = 3 biologically independent samples. One-tailed Student t -tests were performed, SPINDOC KO vector, BRCA1, P = 0.0.0046, 0.0043; NRF1: ** P = 0.0025; P130: ** P = 0.0095; hNIS: * P = 0.0321. SPINDOC KO-GFP-S P INDOC, RAD51: ** P = 0.0018; NRF1: ** P = 0.0038; NF-κB1: ** P = 0.0042; P130: ** P = 0.0013, 0.0080; *** P < 0.001, **** P < 0.0001, NS: nonsignificant.

Article Snippet: Media was changed 4~6 h after transfected cells and cells were cultured for another 24 h. Next, cells were treated with (or without) PJ34 (Selleckchem, Cat# S7300) 10μM an hour and then changed the media with 200 ng/mL Hoechst33342 (Thermo Scientific, Cat# 62249) for 30 mins prior to laser-stripe microirradiation and imaging.

Techniques: Live Cell Imaging, Imaging, Irradiation, Mutagenesis, Transfection, Plasmid Preparation, Incubation, One-tailed Test, Western Blot, In Vitro, Purification, Recombinant, Quantitative RT-PCR, Control

Journal: The Journal of Biological Chemistry

Article Title: Tubulin hyperacetylation drives HMGB1 nuclear exit via the ROS-PARP1 axis, leading to rotenone-induced G2/M arrest

doi: 10.1016/j.jbc.2025.110695

Figure Lengend Snippet: Rotenone triggers time-dependent HMGB1 nuclear exit coupled with increased PARylation. A , representative confocal images of time-dependent HMGB1 ( green ) localization in SH-SY5Y cells treated with rotenone (5μM) or DMSO control. DAPI ( blue ) indicates nuclear staining (scale bar = 10 μm). B , quantification of SH-SY5Y cells with cytoplasmic enrichment of HMGB1 shown in ( A ). Nucleus was defined by area of DAPI. Cytosolic HMGB1 was calculated by subtracting nuclear region of interest (ROI) from the area of green channel. n = 100 cells for each quantification. C , immunoblot analysis of HMGB1 protein levels in the nuclear and cytosolic fraction of SH-SY5Y cells treated with rotenone for 2, 4, 6 and 24 h and compared with DMSO treated controls. GAPDH was used as cytosolic loading control and lamin A/C was used as a nuclear loading control ( left panel ). The nuclear to cytoplasmic ratio was quantified using the lower band obtained in the cytoplasmic fraction of the gels ( right panel ). D , co-immunoprecipitation analysis was performed to assess the PARylation status of HMGB1 in whole-cell extracts following 24 h of rotenone treatment. Cells were immunoprecipitated with an anti-HMGB1 antibody, and PAR ( upper panel ) and HMGB1 ( middle panel ; low and high exposures) levels were analyzed. Additionally, cells were immunoprecipitated with an anti-PAR antibody, and HMGB1 levels ( lower panel ) were examined. E , immunoblot analysis of HMGB1 protein levels in the nuclear and cytosolic fraction of cells treated with rotenone for 24 h in the presence or absence of the PARP inhibitor PJ34 (50μM). GAPDH was used as cytosolic loading control and lamin A/C was used as a nuclear loading control ( left panel ). The nuclear to cytoplasmic ratio was quantified using the lower band obtained in the cytoplasmic fraction of the gels ( right panel ). F , representative confocal images of HMGB1 ( green ) and DAPI (blue) in SH-SY5Y cells treated with rotenone or DMSO control in the presence of PJ34. DAPI ( blue ) indicates nuclear staining (scale bar = 10 μm). G , quantification of cells with cytoplasmic enrichment of HMGB1. n = 100 cells for each quantification. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparison test (∗∗∗ p < 0.001; n = 3 biological replicates).

Article Snippet: PARP inhibitor PJ34 (#HY-13688A; Medchemexpress) SIRT1 inhibitor EX527 (#ab141506; Abcam) was purchased from Abcam.

Techniques: Control, Staining, Western Blot, Immunoprecipitation, Comparison

Journal: The Journal of Biological Chemistry

Article Title: Tubulin hyperacetylation drives HMGB1 nuclear exit via the ROS-PARP1 axis, leading to rotenone-induced G2/M arrest

doi: 10.1016/j.jbc.2025.110695

Figure Lengend Snippet: PARylation primes HMGB1 for acetylation following rotenone treatment that is associated with decreased SIRT1 activity. A , graphical representation of post-translational modification induced by PARP1 that modulates SIRT1-HMGB1 interaction during ongoing DNA damage. B , determination of SIRT1 activity by fluorometric assay in SH-SY5Y cells following rotenone treatment in the presence or absence of PJ34 and compared with DMSO-treated controls. Results are expressed in arbitrary fluorescence units. C , immunoprecipitation analysis with anti-HMGB1 antibody to check the acetylation status of HMGB1 in the whole cell extracts of cells following rotenone treatment for 24 h and probing with pan-acetyl antibody. D and E , representative confocal images of cells stained with SIRT1 ( red ), HMGB1 ( green ) and DAPI ( blue ) following 4 ( D ) or 12 ( E ) h rotenone treatment. (scale bar = 10 μm). F , quantification of SIRT1 intensity at 12 h post rotenone treatment. (n = 100 cells for each quantification). G , representative confocal images of cells stained with SIRT1 (red), HMGB1 ( green ), and DAPI ( blue ) following 24 h rotenone treatment in the presence or absence of PJ34 or a combination of PJ34 and the SIRT1 inhibitor EX527. (scale bar = 10 μm). H , MTT assay of cells treated with rotenone for 24 h in the presence or absence of PJ34 or a combination of PJ34 and EX527. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparison test (∗∗ p < 0.01; ∗∗∗ p < 0.001; n = 3 biological replicates).

Article Snippet: PARP inhibitor PJ34 (#HY-13688A; Medchemexpress) SIRT1 inhibitor EX527 (#ab141506; Abcam) was purchased from Abcam.

Techniques: Activity Assay, Modification, Fluorescence, Immunoprecipitation, Staining, MTT Assay, Comparison

Journal: bioRxiv

Article Title: Parthanatos-inducing zinc agent C010DS-Zn elicits anti-tumor immune responses involving T cells and macrophages in vivo

doi: 10.1101/2021.03.18.433812

Figure Lengend Snippet: Basic substance information on C010DS-Zn, and its control substances C005D-Zn, zinc sulfate (ZnSO 4 ), and “Free Zn-Pyr (7:1)” (the mixture of zinc sulfate and sodium pyrithione at zinc-to-pyrithione molar ratio of 7-to-1). (A) Molecular structure of C010DS, 34 kDa (M w ) γPGA polymer each conjugated with 1 Cy5.5 label, 2 PEG-folate sidechains, 2 PEG-cRGDfK sidechains, and 10 PEG-S-S-pyrithione sidechains. C010DS-Zn preparation is described in the Methods. (B) Molecular structure of C005D, 34 kDa (M w ) γPGA polymer each conjugated with 1 Cy5.5 label, 3 PEG-folate sidechains, and 3 PEG-cRGDfK sidechains. C005D-Zn preparation is described in the Methods. (C) Comparative in vitro cytotoxicity evaluation on the tested compounds using LDH release assays after 24h treatments against 4T1 cells. (D) Results of the in vitro time-resolved apoptosis-necrosis flow cytometry assay at fixed doses against 4T1 cells using A5 and PI labels. (E) Results of the in vitro dose-resolved apoptosis-necrosis flow cytometry assay at 3h time point against 4T1 cells using A5 and PI labels. (F) In vitro PAR-ELISA assay results on the C010DS-Zn treated 4T1 cells with or without the PARP inhibitor PJ34. * p <0.05. ¶ Indicated group displayed significantly higher PAR signal than the Control, 2.5 μM Zn, or 5 μM Zn C010DS-Zn treatment groups at p <0.05 with or without PJ34 co-treatment.

Article Snippet: PJ34 (Santacruz Biotech Cat# sc-204161A) stock solution was prepared at 2mM concentration in water.

Techniques: Control, Polymer, In Vitro, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Parthanatos-inducing zinc agent C010DS-Zn elicits anti-tumor immune responses involving T cells and macrophages in vivo

doi: 10.1101/2021.03.18.433812

Figure Lengend Snippet: In vitro attenuation of 1.5h C010DS-Zn treatment effects against 4T1 including nuclear AIF translocation, DNA fragmentation, and cell viability loss by PARP inhibitor PJ34. (A) Representative confocal fluorescence images of the 4T1 cells treated for 1.5h with vehicle, ZnSO4, C005D-Zn, or C010DS-Zn at different concentrations, labelled with Hoechst (blue: adherent cell count), TUNEL (green: DNA breaks), and anti-AIF (red), with or without the PARP inhibitor PJ34 co-incubation. Bar=100μm. (B) Quantitative analysis of the fluorescence images for cell viability, nuclear AIF translocation, and nuclear TUNEL intensity using CirAvgInten function. 3 imaging fields quantified for each group (n=3). * p <0.05. ** p <0.005. *** p <0.0005. ¶Indicated group’s average value is significantly greater than those of all other treatment groups minimally at p <0.05. C010DS-Zn at 60 μM Zn treatment group resulted in complete cell loss, and hence its quantitative imaging analyses could not be performed.

Article Snippet: PJ34 (Santacruz Biotech Cat# sc-204161A) stock solution was prepared at 2mM concentration in water.

Techniques: In Vitro, Translocation Assay, Fluorescence, Cell Counting, TUNEL Assay, Incubation, Imaging

Journal: Antioxidants

Article Title: Neurotoxic Effect of Myricitrin in Copper-Induced Oxidative Stress Is Mediated by Increased Intracellular Ca 2+ Levels and ROS/p53/p38 Axis

doi: 10.3390/antiox14010046

Figure Lengend Snippet: Effects of specific inhibitors and modulators of intracellular signalling pathways on the neurotoxic effect of myricitrin. Cells were treated simultaneously with 0.5 mM CuSO 4 and 20 µg/mL myricitrin in combination with the following inhibitors: pifithrin-α (p53 inhibitor) ( A ), PJ34 (PARP inhibitor) ( B ), wortmannin (PI3K/Akt inhibitor) ( C ), U0126 (ERK1/2 inhibitor) ( D ), SB203580 (p38 inhibitor) ( E ), SP600125 (JNK inhibitor) ( F ), BAPTA-AM (intracellular calcium chelator) ( G ), MK-801 (NMDA receptor channel blocker) ( H ), nifedipine L-type calcium channel inhibitor) ( I ), and leupeptin (calpain inhibitor) ( J ). Inhibitors were applied 1 h prior to and during the 24 h treatment with copper and myricitrin. Data are presented as means ± SD from 3 to 5 independent experiments performed in triplicate. * p < 0.05, ** p < 0.01, and **** p < 0.0001 vs. copper + myricitrin treatment (one-way ANOVA followed by post hoc Tukey’s test).

Article Snippet: MK-801 hydrogen maleate, pifithrin-α, SP600125, SB203580, and PARP inhibitor VIII (PJ34) were purchased from Alfa Aesar (Ward Hill, MA, USA).

Techniques:

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