Journal: Biomedicines

Article Title: Valemetostat–SAHA-Driven Acetylation of p53 via SET/TAF-Iβ Displacement and p300 Activation Modulates Cell Cycle Regulators in Pancreatic Cancer Cells

doi: 10.3390/biomedicines13092279

Figure Lengend Snippet: p53 acetylation in response to SAHA/DS treatment correlates with changes in its interaction with p300 acetyltransferase and the cochaperone SET/TAF-Iβ. ( A ) Acetylation of p300 was investigated by immunoprecipitation with anti-acetyl-lysine antibody and blotting with an anti-p300 antibody in PaCa44 cancer cells untreated (CT) or undergoing SAHA or SAHA/DS treatment. Beads are shown as loading control. ( B ) Acetyl-p53 (ac p53) as evaluated by Western blotting in PaCa44 and PT45 cells pre-treated (+) or not (−) by p300 inhibitor (p300 inh) and then treated with SAHA/DS or left untreated (CT). β-Actin and GAPDH represent loading controls. ( C ) p21 expression level as evaluated in PaCa44 cancer cells pre-treated (+) or not (−) with p300 inhibitor and exposed to SAHA, SAHA/DS, and DS or untreated control (CT). GAPDH was used as loading control. ( D ) Interaction of mutp53 with p300 and SET/TAF1β investigated by immunoprecipitation in PaCa44 cells treated with SAHA, SAHA/DS, and DS. P53 is shown as loading control. Histograms represent mean plus SD of densitometric analysis from three experiments and expressed as ratio between ac p300/beads and p300/β-Actin ( A ), ac p53/β-Actin and ac p53/GAPDH ( B ), p21/GAPDH ( C ), and p300/p53 and SET/TAF1/p53 ( D ). p -value * < 0.05; ** < 0.01; *** < 0.001; and **** < 0.0001 as calculated by ANOVA test.

Article Snippet: In some experiments, cells were pre-treated for 60 min with the p300 inhibitor A-485 (p300 inh) (100 μM) (Selleckchem, Cologne, Germany, cat. S8740) before exposure to SAHA/DS.

Techniques: Immunoprecipitation, Control, Western Blot, Expressing

Journal: Advanced Science

Article Title: HPV E6/E7‐Induced Acetylation of a Peptide Encoded by a Long Non‐Coding RNA Inhibits Ferroptosis to Promote the Malignancy of Cervical Cancer

doi: 10.1002/advs.202414018

Figure Lengend Snippet: Histone acetyltransferase CBP/p300 promotes H3K27 acetylation of lncTUBA3FP enhancer. A) Predicted H3K4Me1, H3K4Me3, and H3K27ac enrichments at lncTUBA3FP enhancer region using the UCSC website. B) ChIP of the enrichment of H3K27ac in lncTUBA3FP enhancer region in E6 or E7 knockdown HeLa cells ( sgHPV E6, sgHPV E7 ) or the control ( Lenti‐V2 ) ( n = 3). C) ChIP‐qPCR analysis of the H3K27ac level in lncTUBA3FP enhancer in C‐33A cells transfected with HPV E6 or E7 construct ( HPV E6, HPV E7 ) or the control ( pcDNA3.1 ) ( n = 3). D) RT‐qPCR analysis of the lncTUBA3FP level in HeLa cells transfected with CBP‐HA ( CBP ), p300‐HA ( p300 ), and their control ( pcDNA3.1 ) ( n = 3). E) H3K27ac level in lncTUBA3FP enhancer in HeLa cells treated as in (D) was detected by ChIP‐qPCR ( n = 3). F) Western blotting analysis of the TUBORF peptide level in HeLa cells treated as in (D). G) HeLa cells were treated with A‐485 (2 or 10 µ m in 24 h) or DMSO as the control. RT‐qPCR analysis was performed to examine the lncTUBA3FP level ( n = 4). H) ChIP analysis of the H3K27ac level in lncTUBA3FP enhancer in HeLa cells treated with A‐485 (10 µ m , 24 h) or DMSO as the control ( n = 3). I) Western blotting analysis of the TUBORF peptide level in HeLa cells treated as in (H). Data were presented with mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001, Student's t ‐test.

Article Snippet: The cells were respectively treated with 20 μg mL −1 Cycloheximide (CHX) (Cell Signal Technology) for different time‐points, proteasome inhibitor MG132 (Selleck) at 10 μ m for 6 h, ferroptosis inducer Erastin (MCE) for 24 h, or CBP/p300 catalytic inhibitor A‐485 at 10 μ m for 24 h. siRNA oligonucleotides targeting HPVs E6/E7 were purchased from GenePharma as previously described.

Techniques: Knockdown, Control, ChIP-qPCR, Transfection, Construct, Quantitative RT-PCR, Western Blot

Journal: Advanced Science

Article Title: HPV E6/E7‐Induced Acetylation of a Peptide Encoded by a Long Non‐Coding RNA Inhibits Ferroptosis to Promote the Malignancy of Cervical Cancer

doi: 10.1002/advs.202414018

Figure Lengend Snippet: Knockdown of ESCO1 or TUBORF enhances paclitaxel‐initiated cell ferroptosis and amplifies the anticancer effect of paclitaxel in cervical cancer. A) ESCO1 knockout cells ( ESCO1 KO #1 ), lncTUBA3FP knockout cells ( lncTUBA3FP KO #1 ), or the control ( WT ) were treated with 5 µ m Erastin, 10 µ m paclitaxel or DMSO for 24 h. Cells were collected for Western blotting analysis of SLC7A11 and GPX4 protein levels. B) Cells treated as in (A) were employed to measure Fe 2+ levels ( n = 3). C) Cells treated as in (A) were used to examine GSH/GSSG levels ( n = 3). D) Cells treated as in (A) were used to measure lipid ROS levels ( n = 3). E) Soft agar colony formation assay of cells treated as in (A). The representative images were photographed two weeks after cell seeding. Magnification, ×100. Scar bars, 40 µm. F) Results were quantified in (E) ( n = 11). G) Representative isolated tumor images from nude mice with paclitaxel or DMSO treatment and injected with ESCO1 knockout ( ESCO1 KO #1 ), lncTUBA3FP knockout ( lncTUBA3FP KO #1 ), and wild type ( WT ) transduced SiHa cells after 28 days were shown ( n = 6). H) Tumor volumes in (G) were measured every 3 days starting from day 7 ( n = 6). I) Tumor weights in (G) were recorded after the isolation of tumors ( n = 6). J) Tissues treated as in (G) were employed to measure Fe 2+ levels, with a portion of each tissue being selected for combined detection ( n = 3). K) Tissues treated as in (G) were used to examine GSH/GSSG levels, with a portion of each tissue being selected for combined detection ( n = 3). L) Tissues treated as in (G) were used to measure lipid ROS levels, with a portion of each tissue being selected for combined detection ( n = 3). M) A schematic working model of HPV‐induced TUBORF acetylation. HPV oncoproteins E6 and E7 upregulate lncTUBA3FP level by histone H3K27ac modification subjected to acetyltransferase CBP/p300, which encodes a peptide named TUBORF. E6 and E7 increase and recruit acetyltransferase ESCO1 to bind to and acetylate TUBORF. Acetylated TUBORF binds to IRGQ protein and promotes its degradation, resulting in inhibition of ferroptosis and enhancement of the malignant proliferation of cervical cancer. Data were presented with mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001, Student's t ‐test.

Article Snippet: The cells were respectively treated with 20 μg mL −1 Cycloheximide (CHX) (Cell Signal Technology) for different time‐points, proteasome inhibitor MG132 (Selleck) at 10 μ m for 6 h, ferroptosis inducer Erastin (MCE) for 24 h, or CBP/p300 catalytic inhibitor A‐485 at 10 μ m for 24 h. siRNA oligonucleotides targeting HPVs E6/E7 were purchased from GenePharma as previously described.

Techniques: Knockdown, Knock-Out, Control, Western Blot, Soft Agar Assay, Isolation, Injection, Modification, Inhibition

Journal: bioRxiv

Article Title: Super-resolution microscopy reveals distinct epigenetic states regulated by estrogen receptor activity

doi: 10.1101/2025.06.16.659976

Figure Lengend Snippet: A) Confocal images of MCF7 cells probed for H3K27ac (magenta) and ERα (green) under estrogen-deprived (ED) and E2-treated (10 nM) conditions, highlighting the resolution limit of conventional microscopy. B) Super-resolution 3D-SIM images of MCF7 cells under ED, E2 (10 nM), Tam (100 nM), and Fulv (100 nM) conditions, demonstrating enhanced proximity between ERα and H3K27ac in E2-treated cells. C) Boxplots of pixel-level correlation coefficients between ERα and H3K27ac signal intensities in H3K27ac domains for WT and Y537S mutant MCF7 cells under ED, E2, Tam, and Fulv conditions. Statistics (one-side student T-test) were provided for E2 compared with other conditions: *P < 0.01, **P < 0.001, ***P < 0.0001. D) Boxplot with overlaid data points showing the t-statistic from each data batch. The t-statistic quantifies the difference in correlation between H3K27ac and ER signal intensity in E2 compared to ED. Positive and negative scores indicate higher and lower correlation, respectively. E) 3D-SIM images of MCF7 cells probed for p300 (magenta) and ERα (green) under ED, E2, Tam, and Fulv conditions. F) Violin plots of colocalization coefficients between p300 and ERα in wild-type and Y537S mutant MCF7 cells under different conditions.

Article Snippet: Treatments included estradiol (E2, Sigma-Aldrich, Cat# E2758, 10 nM), tamoxifen (Tam, Sigma-Aldrich, Cat# T5648, 100 nM), fulvestrant (Fulv, Sigma-Aldrich, Cat# I4409, 100 nM), and the p300 inhibitor A-485 (MedChemExpress, Cat# HY-100231, 80 nM).

Techniques: Microscopy, Mutagenesis

Journal: bioRxiv

Article Title: Super-resolution microscopy reveals distinct epigenetic states regulated by estrogen receptor activity

doi: 10.1101/2025.06.16.659976

Figure Lengend Snippet: A) 2-color 3D STORM images of MCF7 cells treated with E2 (10 nM), Tam (100 nM), and Fulv (100 nM). Tam and Fulv treatments show more “closed” H3K27ac and ERα structures compared to the “open” structures in E2-treated cells. B, C) 3-D STORM images of MCF7 cells under control siRNA, NCOA3 siRNA, and p300 inhibitor A485 (80 nM) conditions. The “open” structures in E2-treated control siRNA cells are lost upon NCOA3 knockdown or p300 inhibition.

Article Snippet: Treatments included estradiol (E2, Sigma-Aldrich, Cat# E2758, 10 nM), tamoxifen (Tam, Sigma-Aldrich, Cat# T5648, 100 nM), fulvestrant (Fulv, Sigma-Aldrich, Cat# I4409, 100 nM), and the p300 inhibitor A-485 (MedChemExpress, Cat# HY-100231, 80 nM).

Techniques: Control, Knockdown, Inhibition

Journal: bioRxiv

Article Title: Super-resolution microscopy reveals distinct epigenetic states regulated by estrogen receptor activity

doi: 10.1101/2025.06.16.659976

Figure Lengend Snippet: A, B) 3D renderings of STORM images from , classifying structures as “open” or “closed” for each condition. B) Rendering of STORM images from , showing “closed” vs. “open” structures in cells lacking NCOA3 and p300 activity. C&D) Structural maps of rendered images from parts A, B, classifying H3K27ac modified chromatin into four volume classes (yellow, green, blue = high volume; purple = low volume). E) Distribution of volume classes across conditions.. E2-treated cells show more high-volume, “open” structures, while Tam, Fulv, NCOA3 siRNA, and A485 treatments shift toward low-volume, “closed” structures. Significance: *P < 0.01, **P < 0.001, ***P < 0.0001. F, G) Violin plots of H3K27ac domain volumes (F) and sphericity (G) per condition. Kruskal-Wallis ANOVA with Mann-Whitney U test and Bonferroni correction. Significance: *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Treatments included estradiol (E2, Sigma-Aldrich, Cat# E2758, 10 nM), tamoxifen (Tam, Sigma-Aldrich, Cat# T5648, 100 nM), fulvestrant (Fulv, Sigma-Aldrich, Cat# I4409, 100 nM), and the p300 inhibitor A-485 (MedChemExpress, Cat# HY-100231, 80 nM).

Techniques: Activity Assay, Modification, MANN-WHITNEY