Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Palladium-catalysed synthesis of small-molecule epigenetic inhibitors as anticancer therapeutics

doi: 10.1080/14756366.2026.2621477

Figure Lengend Snippet: Design Strategy & Synthetic Route of Novel Benzofuran Derivatives as Potent LSD1 Inhibitors (the figure was drawn by the authors using chemdraw software).

Article Snippet: Duan et al. designed 5-cyano-3-phenylindole-based LSD1/HDAC dual inhibitors for colorectal cancer, inspired by the Novartis LSD1 inhibitor ( 350 ).

Techniques: Software

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Palladium-catalysed synthesis of small-molecule epigenetic inhibitors as anticancer therapeutics

doi: 10.1080/14756366.2026.2621477

Figure Lengend Snippet: Design Strategy & Synthetic Route of Novel Indole Derivatives as LSD1 Inhibitors (the figure was drawn by the authors using chemdraw software).

Article Snippet: Duan et al. designed 5-cyano-3-phenylindole-based LSD1/HDAC dual inhibitors for colorectal cancer, inspired by the Novartis LSD1 inhibitor ( 350 ).

Techniques: Software

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Palladium-catalysed synthesis of small-molecule epigenetic inhibitors as anticancer therapeutics

doi: 10.1080/14756366.2026.2621477

Figure Lengend Snippet: Design Strategy & Synthetic Route of Dual EZH2 & LSD1 Inhibitor (the figure was drawn by the authors using chemdraw software).

Article Snippet: Duan et al. designed 5-cyano-3-phenylindole-based LSD1/HDAC dual inhibitors for colorectal cancer, inspired by the Novartis LSD1 inhibitor ( 350 ).

Techniques: Software

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Palladium-catalysed synthesis of small-molecule epigenetic inhibitors as anticancer therapeutics

doi: 10.1080/14756366.2026.2621477

Figure Lengend Snippet: Design Strategy & Synthetic Route of 2‑Aryl-4-aminoquinazolin-Based LSD1 Inhibitors (the figure was drawn by the authors using chemdraw software).

Article Snippet: Duan et al. designed 5-cyano-3-phenylindole-based LSD1/HDAC dual inhibitors for colorectal cancer, inspired by the Novartis LSD1 inhibitor ( 350 ).

Techniques: Software

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Palladium-catalysed synthesis of small-molecule epigenetic inhibitors as anticancer therapeutics

doi: 10.1080/14756366.2026.2621477

Figure Lengend Snippet: Design Strategy & Synthetic Route of Novel 1H‑Pyrrolo[2,3‑c]pyridin Derivatives Based Lysine Specific Demethylase 1 Inhibitors (the figure was drawn by the authors using chemdraw software).

Article Snippet: Duan et al. designed 5-cyano-3-phenylindole-based LSD1/HDAC dual inhibitors for colorectal cancer, inspired by the Novartis LSD1 inhibitor ( 350 ).

Techniques: Software

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Palladium-catalysed synthesis of small-molecule epigenetic inhibitors as anticancer therapeutics

doi: 10.1080/14756366.2026.2621477

Figure Lengend Snippet: Design Strategy & Synthetic Route of Dual LSD1 and HDAC Inhibitor (the figure was drawn by the authors using chemdraw software).

Article Snippet: Duan et al. designed 5-cyano-3-phenylindole-based LSD1/HDAC dual inhibitors for colorectal cancer, inspired by the Novartis LSD1 inhibitor ( 350 ).

Techniques: Software

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Palladium-catalysed synthesis of small-molecule epigenetic inhibitors as anticancer therapeutics

doi: 10.1080/14756366.2026.2621477

Figure Lengend Snippet: Design Strategy & Synthetic Route of Novel acridine-based LSD1 inhibitors (the figure was drawn by the authors using chemdraw software).

Article Snippet: Duan et al. designed 5-cyano-3-phenylindole-based LSD1/HDAC dual inhibitors for colorectal cancer, inspired by the Novartis LSD1 inhibitor ( 350 ).

Techniques: Software

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Palladium-catalysed synthesis of small-molecule epigenetic inhibitors as anticancer therapeutics

doi: 10.1080/14756366.2026.2621477

Figure Lengend Snippet: Design Strategy & Synthetic Route of Dual LSD1 & HDAC Inhibitor (the figure was drawn by the authors using chemdraw software).

Article Snippet: Duan et al. designed 5-cyano-3-phenylindole-based LSD1/HDAC dual inhibitors for colorectal cancer, inspired by the Novartis LSD1 inhibitor ( 350 ).

Techniques: Software

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Palladium-catalysed synthesis of small-molecule epigenetic inhibitors as anticancer therapeutics

doi: 10.1080/14756366.2026.2621477

Figure Lengend Snippet: Design Strategy & Synthetic Route of Novel 5‑Cyano-3-phenylindole-Based LSD1/HDAC Dual Inhibitors (the figure was drawn by the authors using chemdraw software).

Article Snippet: Duan et al. designed 5-cyano-3-phenylindole-based LSD1/HDAC dual inhibitors for colorectal cancer, inspired by the Novartis LSD1 inhibitor ( 350 ).

Techniques: Software

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Palladium-catalysed synthesis of small-molecule epigenetic inhibitors as anticancer therapeutics

doi: 10.1080/14756366.2026.2621477

Figure Lengend Snippet: Design Strategy & Synthetic Route of CC-90011 (328): A Lysine Specific Demethylase 1 Inhibitors (the figure was drawn by the authors using chemdraw software).

Article Snippet: Duan et al. designed 5-cyano-3-phenylindole-based LSD1/HDAC dual inhibitors for colorectal cancer, inspired by the Novartis LSD1 inhibitor ( 350 ).

Techniques: Software

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Schematic of the LSD1/ZNF217/CoREST complex, which can erase H3K4 methyl and/or H3K9 acetyl marks. ( B-D ) Immunoblot analysis of whole cell lysates (WCL) from Cas9+ Akata cells expressing control, LSD1 (B), ZNF217 (C), or CoREST (D) sgRNAs for immediate early lytic BZLF1, early BMRF1 and late p18 expression. ( E ) Fluorescence-activated cell sorting (FACS) analysis of PM gp350 levels on Akata cells expressing control, LSD1, ZNF217, or CoREST sgRNAs. ( F ) Mean fluorescence intensity (MFI) ± standard deviation (SD) gp350 PM values from n=3 independent replicates. ( G-H ) Intracellular (G) or extracellular (H) qPCR analysis of EBV genome copy number in Akata cells expressing control, LSD1, ZNF217, or CoREST sgRNA. Shown are mean ± SD values from n=3 replicates. P-values indicate significance of differences relative to control sgRNA cell values. ( I ) Immunoblot analysis of WCL from Akata cells expressing control, LSD1, ZNF217, or CoREST sgRNA. Blots are representative of n=3 replicates. * p <0.05, ** p <0.01, *** p < 0.001 by Student’s t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Western Blot, Expressing, Control, Fluorescence, FACS, Standard Deviation

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Immunoblot analysis of WCL from P3HR-1 or MUTU I Burkitt cells treated with the indicated corin concentrations for two days. (B) FACS analysis of PM gp350 levels in corin-treated P3HR-1 or MUTU I cells. (C) Immunoblot analysis of WCL from EBV+ KEM III, Jijoye, Farage, AGS, YCCEL1, C666-1 cells treated with corin (0, 1, 2, or 5 μM) for two days. ( D ) Immunoblot analysis of WCL from EBV+/KSHV+ JSC-1 primary effusion lymphoma cells treated with corin (0, 1, 2, or 5 μM) for two days. ( E ) Workflow of corin and ganciclovir (GCV) treatment. Cells were seeded into fresh media on days of corin treatment. GCV (10 μg/ml) was added twice daily where indicated. ( F ) EBV+ Burkitt B-cells were treated as described in (E), with the following corin concentrations: P3HR-1 (0.5 μM), MUTU I (0.5 μM), Rael (0.25 μM) vs vehicle. Shown are mean ± SD live cell number relative to DMSO-treated controls on day 6 from n=3 replicates. Values of DMSO treated control cells were normalized to 1. * p <0.05, *** p < 0.001. ( G ) MUTU I murine xenograft experiment workflow. Two weeks post MUTU I Burkitt xenograft implantation, mice were treated with vehicle or corin (30 mg/ml), as indicated. ( H ) Immunohistochemical analysis of BMRF1 expression in xenograft tumors following treatment with vehicle vs. corin as in Figure 3G . Scale bar = 100 μm. ( I ) Dosing strategy for analysis of corin, GCV or corin+GCV effects on murine MUTU I xenograft tumor size. Once xenograft tumors reached a volume of 50mm 3 , n=6 or 7 mice were treated with vehicle, corin (30mg/kg), GCV (50mg/kg) or corin (30mg/kg) + GCV (50mg/kg). ( J ) Area under the curve (AUC) analysis of tumor size up to 21 days post the initial treatment. AUC integrates tumor size measurements per group across all timepoints. All blots shown are representative images of n=3 replicates. * p <0.05, *** p < 0.001 by Student’s t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Inhibition, Western Blot, Control, Immunohistochemical staining, Expressing

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Chromatin immunoprecipitation sequencing (ChIP-seq) analysis of EBV genome-wide ZNF217 and LSD1 occupancy in latently infected Akata cells. Shown are input, ZNF217 and LSD1 ChIP-seq tracks from n=2 independent replicates, with track heights set to 5,000 (5K). Shown at top are EBV gene annotations of regions with ZNF217 and/or LSD1 ChIP-seq peaks, and as a control, the BXLF1 promoter region that lacks ZNF217 or LSD1 peaks. ( B ) Mean ± SD % of input values from n=3 ChIP-qPCR replicates of LSD1 occupancy at oriLyt , the BZLF1 promoter (BZLF1p) or the BXLF1 in Cas9+ Akata cells with control (blue) versus ZNF217 sgRNA (orange) sgRNAs. Shown at right are negative control IgG ChIP-qPCR values. ( C ) Mean ± SD % of input values from n=3 ChIP-qPCR replicates of ZNF217 occupancy as in ( B ). Locations of the BLLF3 promoter (BLLF3p) and BSLF1 promoter (BSLF1p) regions, which exhibited ZNF217 but not LSD1 ChIP-seq peaks. ( D ) Immunoblot analysis of WCL from Cas9+ Akata cells that stably expressed control GFP or wildtype versus deletion mutant ZNF217 CRISPR resistant cDNAs and that co-expressed negative control or ZNF217-targeting sgRNA to deplete endogenous ZNF217. The schematic at bottom depicts the ZNF217 N-terminal CoREST binding domain (aa 1–272), DNA binding domain (residues 471–526) and CTBP binding domain (aa 686–760). Blots are representative of n=3 replicates. ( E ) FACS analysis of PM gp350 abundances in Cas9+ Akata cells with stable control GFP or CRISPR resistant ZNF217 cDNAs that co-expressed control or ZNF217 sgRNA to deplete endogenous ZNF217 (as in panel D ). ( F ) ChIP-qPCR analysis of H3K4 mono (H3K4me1), di (H3K4me2) and tri (H3K4me3) methylation levels in Cas9+ Akata expressing control sgRNA or either of two independent screen hit sgRNAs against LSD1, ZNF217 or CoREST. Mean ± SD % of input values from n=3 replicates ChIP-qPCR replicates of H3K4me1, 2 or 3 levels at oriLyt L in Cas9+ Akata cells that expressed the indicted sgRNAs. Significance values in panel F refer to comparisons between cells with the indicated vs control sgRNAs. NS, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001 by Student’s t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Methylation, ChIP-sequencing, Genome Wide, Infection, Control, ChIP-qPCR, Negative Control, Western Blot, Stable Transfection, Mutagenesis, CRISPR, Binding Assay, Expressing

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Schematic diagram highlighting chromatin conformation capture (3C) assay BZLF1p anchor primer (red box) and 12 test (T) primers locations along the linear EBV genome, including at oriLyt R (primer T6). For reference, EBV genomic terminal repeat ( TR ) and origin of plasmid replication ( oriP ) are shown. ( B ) 3C assay analysis of DNA looping between the BZLF1 anchor and 12 test primer regions. Shown are the mean ± SD 3C assay signals relative to EBV BACmid negative control from Akata cells expressing control, LSD1, ZNF217, or CoREST sgRNAs. ( C ) Mean ± SD 3C assay signals from Akata cells 24 hours post-treatment with vehicle, C12 (2.5 μM), C12 (2.5 μM) + NaB (0.5 mM), or corin (2.5 μM). * p < 0.05; ** p < 0.01; *** p < 0.001 by Student’s t-test. ( D ) Schematic model. In latency, LSD1/ZNF217/CoREST/HDAC complexes co-occupy EBV oriLyt enhancer regions, where LSD1 and HDAC erase activating H3K4 methylation and H3K4 acetylation marks, respectively. Perturbation of LSD1 and HDAC activity enables KMT2D to deposit activating H3K4 epigenetic marks at both oriLyt enhancers and supports their looping to the immediate early BZLF1 promoter. Newly synthesized BZLF1 drives the lytic cycle by binding to early gene promoters and also to oriLyt enhancers to increase their strength, which further upregulates BZLF1 in a positive feedback loop.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Plasmid Preparation, Negative Control, Expressing, Control, Methylation, Activity Assay, Synthesized, Binding Assay

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) STRING interaction network analysis 94 of LSD1/ZNF217/CoREST complex and its associated factors. Shown are edges, which depict protein-protein associations, and confidence scores, which depict the estimated likelihood, based on supporting evidence, that the predicted interaction is biologically meaningful, specific and reproducible. ( B ) Immunoblot analysis of WCL from Akata cells expressing control or CTBP1 targeting sgRNA. ( C ) FACS analysis of PM gp350 abundance in Akata cells expressing control or independent Brunello library CTBP1 targeting sgRNAs. ( D ) Immunoblot analysis of WCL from Akata cells expressing control or BCL6 targeting sgRNA. ( E ) FACS analysis of PM gp350 levels in Akata cells expressing control or BCL6 targeting sgRNA. ( F ) Immunoblot analysis of WCL from Akata cells expressing control or independent Brunello PHF12 sgRNAs. ( G ) FACS analysis of PM gp350 levels in Akata cells expressing control or independent Brunello PHF12 targeting sgRNAs. ( H ) Mean ± SD intracellular EBV genome copy number from n=3 replicates of Akata cells expressing control, CTBP1, BCL6, or PHF12 sgRNAs. Blots shown are representative of n=3 replicates. ** p < 0.01, *** p < 0.001 by Student’s t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Western Blot, Expressing, Control

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) WCL of P3HR-1 cells were subjected to immunoprecipitation using control IgG or anti-LSD1 antibody, followed by immunoblot analysis with 1% input for the indicated proteins. ( B ) Left, LSD1, ZNF217 and CoREST relative protein abundances in P3HR-1 ZHT/RHT cells uninduced or induced for lytic replication by 4-HT for the indicated times (right) or in EBV-negative (EBV−) versus EBV-positive (EBV+) Akata mock induced or induced for lytic replication by anti-IgG crosslinking for 48 hours, using data from Ersing et al. 47 . ( C-E ) FACS analysis of PM gp350 abundances in P3HR-1 cells expressing control, LSD1 (C), ZNF217 (D) or CoREST (E) targeting sgRNAs. ( F ) Immunoblot analysis of WCL from EBV super-infected AGS or EBV+ SNU719 gastric carcinoma cells expressing control or ZNF217 sgRNAs. ( G ) Immunoblot analysis of WCL from EBV+ or EBV− Akata cells expressing control or CoREST sgRNA. Blots are representative of n=3 independent replicates.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Immunoprecipitation, Control, Western Blot, Expressing, Infection

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Immunoblot analysis of WCL from latency I EB3 or Rael Burkitt cells, latency III GM15892 LCLs or KEM III LCLs, EBV+ AGS gastric carcinoma and C666-1 nasopharyngeal carcinoma cells treated with C12 (0,1,2 or 5 μM) for 48 hours. ( B ) Immunoblot analysis of WCL from MUTU I that were treated with the indicated concentration of C12 for 6 or 24 hours and then grown in C12-free RPMI for the remainder of 48 hours. ( C ) Immunoblot analysis of WCL from KEM I cells treated with C12 (0, 1, 2, 5, or, 10 μM) alone or in combination with NaB (0.5 mM) for 48 hours. ( D-E ) Immunoblot analysis of WCL (D) and FACS analysis of PM gp350 (E) from P3HR-1 cells treated with C12 (0, 1, 2, 5, or, 10 μM) alone or in combination with NaB (0.5 mM) for 48 hours. ( F-G ) Immunoblot analysis of WCL (F) and FACS analysis of PM gp350 levels (G) from MUTU I cells treated with C12 (0, 1, 2, 5, or, 10 μM) alone or in combination with NaB (0.5 mM) for 48 hours. Shown in the top right of the panels in (E) and (G) are %gp350+ cells. Blots are representative of n=3 independent replicates.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Inhibition, Western Blot, Concentration Assay

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Schematic diagram depicting bifunctional small molecule antagonist corin inhibition of LSD1 and HDAC activity. ( B ) Immunoblot analysis of WCL from P3HR-1, Akata or MUTU I cells that were treated with the indicated concentration of corin for 6 hours and then grown in corin-free RPMI for the remainder of 48 hours. ( C ) Mean ± SD intracellular EBV genome copy number from n=3 replicates of P3HR-1 or MUTU I cells treated with corin for 48 hours. ( D ) Immunoblot analysis of WCL from Akata cells with stable GFP versus MYC cDNA overexpression that were treated with corin for 48 hours, as indicated. ( E ) Immunoblot analysis of WCL from control or BZLF1 knockout (KO) Akata cells treated with corin for 48 hours, suggesting that corin requires BZLF1 to induce early and late protein expression. ( F ) Immunoblot analysis of WCL from Akata, KEM I, EB3, Rael Burkitt cells, GM15892 LCLs or SNU719 gastric carcinoma cells treated with corin for 48 hours. ( G ) Immunoblot analysis of WCL from EBV+/KSHV+ BC-1 or KSHV+ BCBL-1 primary effusion lymphoma cells treated with corin for 48 hours. ( H ) KSHV+ iSLK.219 epithelial cells with conditional doxycycline (Dox) inducible KSHV immediate early RTA expression was treated with DMSO, Dox (0.5 ug/ml) or corin (5 μM) for 24 hours, and then maintained in growth media without Dox or corin. 48 hours later, immunofluorescence analysis was performed for GFP (stably expressed from the KSHV genome) versus for red fluorescence protein (RFP, controlled by lytic PAN RNA promoter, which reports KSHV lytic gene expression). Scale bar = 650 μm. Blots and immunofluorescence images are representative of n=3 independent replicates. ** p <0.01, *** p < 0.001 by Student’s t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Inhibition, Activity Assay, Western Blot, Concentration Assay, Over Expression, Control, Knock-Out, Expressing, Immunofluorescence, Stable Transfection, Fluorescence, Gene Expression

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Corin and ganciclovir co-treatment effects on EBV-negative Burkitt and EBV+ LCL cells. Shown are mean ± SD fold changes of live cell numbers, relative to DMSO-treated controls, from n=3 replicates on day 6 of treatment of EBV-negative MUTU I (left) and of EBV+ GM15892 LCLs (right). Cells were treated as described in Fig. 3E with DMSO or corin (0.5 μM). ( B ) Corin and ganciclovir co-treatment effects on KEM III LCLs. KEM III LCLs were treated with a single dose on day 1 of or with multiple drug doses as described in Fig. 3E . Shown are mean ± SD fold changes of live cell numbers, relative to DMSO-treated controls, from n=3 replicates on day 6 of treatment. ( C ) P3HR-1 Burkitt cells were treated with a single dose on day 1 or with multiple doses of DMSO vehicle, LSD1 inhibitor C12 (0.5 μM), the HDAC inhibitor NaB (0.1 mM), C12 + NaB or with the dual LSD1/HDAC inhibitor corin (0.5 μM), using the regimen described in Fig. 3E . Shown are mean ± SD fold changes of live cell numbers, relative to DMSO-treated controls, from n=3 replicates on day 6 of treatment. ( D ) Immunoblot analysis of WCL prepared from MUTU I xenografts harvested from mice treated with DMSO vehicle vs corin as described in Fig. 3G . ( E ) Immunohistochemical analysis of BZLF1 expression in MUTU I xenografts harvested from mice treated with DMSO vs corin, as described in Fig. 3G . Scale bar = 100 μm. ( F ) Quantification of BZLF1+ (left panel) versus BMRF1+ (right panel) cell numbers in MUTUI xenograft tumors as in Fig. 3H and Extended Data Fig. 6E . Shown are mean ± SEM numbers of BZLF1+ or BMRF1+ cells from four randomly selected fields per mouse, quantitated by Image J using the Cell Counter plugin. ( G ) qPCR analysis of BZLF1, early BMRF1 and late BLLF1 (encodes gp350) mRNA abundances in xenograft tumors from tumors shown in Fig. 3H and Extended Data 6E . ( H ) qPCR defined intracellular EBV genome copy number from the xenograft tumors shown in Fig. 3H . Box heights indicate mean qPCR values. ( I ) MUTU I xenograft tumor sizes from the vehicle control, corin, GCV or corin and GCV treated mice shown in Fig. 3J . Significance was determined by cross-comparison of mean tumor sizes from dual drug treated versus vehicle or single drug treated mice. Blots shown are representative images of n=3 replicates. * p <0.05, ** p <0.01, *** p < 0.001, NS, not significant by Students t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Transformation Assay, In Vivo, Western Blot, Immunohistochemical staining, Expressing, Control, Comparison

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) ChIP-qPCR analysis of CoREST occupancy at oriLyt versus BZLF1p and BXLF1 regions in Akata control versus ZNF217 KO cells. Mean ± SD percentages of input values from n=3 replicates are shown. ( B ) ZNF217 mutant validation. Immunoblot analysis of WCL vs α-V5 epitope tagged control GFP, wildtype or deletion mutant ZNF217 complexes immunopurified from Cas9+ Akata cells that expressed sgRNA against endogenous ZNF217. Red asterisks indicate V5-tagged ZNF217 bands. ( C-D ) ChIP-qPCR analysis of LSD1 ( C ) or CoREST ( D ) occupancies at the BZLF1 promoter or oriLyt L in Cas9+ Akata cells with stable wildtype or DNA binding domain deletion (△471–526) ZNF217 cDNAs and that also expressed negative control or ZNF217 targeting sgRNA. Mean ± SD percentages of input values from n=3 replicates are shown. ( E ) Immunoblot analysis of WCL from Akata cells expressing V5-tagged GFP or ZNF217 cDNAs, with or without α-IgG (10 μg/ml) stimulation for 24 hours ( F ) ChIP-qPCR analysis of BZLF1 promoter or oriLyt LSD1 or CoREST occupancy in Akata cells with stable expression of control GFP or ZNF217 cDNAs. Mean ± SD percentages of input values from n=3 replicates are shown. ( G ) Immunoblot analysis of WCL from Cas9+ Akata cells that expressed negative control sgRNA, an independent sgRNA targeting the oriLyt R or L regions with the highest ChIP-seq defined ZNF217 occupancy (sgRNA #1–4 against oriLyt R and #6–9 against oriLyt L ), or positive control sgRNAs targeting oriLyt R vs oriLyt L region MYC-occupied E-Box sites critical for EBV latency 19 (sgRNA #5 and #10, respectively). ( H ) ChIP-qPCR analysis of ZNF217 (left) or H3K4me1 (right) levels in negative control vs oriLyt region expressing Akata cells. Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. All blots shown are representative images of n=3 replicates. * p <0.05, ** p <0.01, *** p < 0.001; NS: not significant as defined by Student’s t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Methylation, ChIP-qPCR, Control, Mutagenesis, Biomarker Discovery, Western Blot, Binding Assay, Negative Control, Expressing, ChIP-sequencing, Positive Control

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A-B ) ChIP-qPCR analysis of H3K4me1, H3K4me2, and H3K4me3 levels in control vs LSD1, ZNF217 or CoREST depleted Akata cells. Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates of Akata cells expressing control, LSD1, ZNF217, or CoREST sgRNAs, using primers specific for oriLyt R ( A ) or the BZLF1 promoter ( B ). ( C ) ChIP-qPCR analysis of H3K4me1 levels in control vs LSD1 depleted Akata cells in the presence of acyclovir (100 μg/ml). Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates of Akata cells expressing control or LSD1 sgRNAs, using primers specific for BZLF1p, BXLF1, or oriLyt regions. ( D ) ChIP-qPCR analysis of H3K4me1 levels in control vs LSD1, ZNF217, or CoREST depleted Akata cells. Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates of Akata cells expressing control or LSD1, ZNF217, or CoREST sgRNAs, using primers specific for the BXLF1 region. ( E ) Mean ± SD ChIP-qPCR % input values from n=3 replicates of Akata cells expressing control or LSD1 sgRNA, using primers specific for the BZLF1 promoter or oriLyt regions. * p <0.05; ** p < 0.01; *** p < 0.001, NS: not significant as defined by Student’s t-test. Significance values in panels A , B , and E refer to comparisons between cells with the indicated vs control sgRNAs.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: ChIP-qPCR, Control, Expressing

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) ChIP-qPCR analysis of LSD1 and ZNF217 occupancy in Akata cells 24 hours after treatment with vehicle, C12 (2.5 μM) or corin (2.5 μM), in the presence of acyclovir (100 μg/ml) to block EBV lytic DNA synthesis. Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. ( B ) ChIP-qPCR analysis of H3K4me1, H3K4me2 and H3K4me3 abundances at the BZLF1 promoter or oriLyt regions in Akata cells 24 hours after treatment with vehicle, C12 (2.5 μM) or corin (2.5 μM). Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. ( C ) ChIP-qPCR analysis of H3K9Ac and H3K27Ac abundances at the BZLF1 promoter or oriLyt regions in Akata cells 24 hours after treatment with vehicle, C12 (2.5 μM) or corin (2.5 μM). Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. ( D ) ChIP-qPCR analysis of H3K9 and H3K27 acetylation levels in Akata cells expressing control versus independent LSD1 targeting sgRNAs. Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. ( E ) ChIP-qPCR analysis of H3K9me2 and H3K9me3 abundances at the BZLF1 promoter or oriLyt regions in Akata cells treated with vehicle control, C12 (2.5 μM) or corin (2.5 μM) for 24 hours. Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. Student’s t-test was performed to cross-compare ChIP-qPCR values from C12 or corin treated cells or LSD1 KO cells with corresponding values from control cells: * p <0.05, ** p <0.01, *** p < 0.001; NS: not significant.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: ChIP-qPCR, Blocking Assay, DNA Synthesis, Expressing, Control