Journal: International Journal of Molecular Sciences

Article Title: The Effect of Liposomal Curcumin as an Anti-Inflammatory Strategy on Lipopolysaccharide e from Porphyromonas gingivalis Treated Endothelial Committed Neural Crest Derived Stem Cells: Morphological and Molecular Mechanisms

doi: 10.3390/ijms22147534

Figure Lengend Snippet: DNMT1 and p300 expression in hPDLSCs and e-hPDLSCs. The localization of DNMT1 and p300 was determined by immunofluorescence in all considered sample groups. Positive cells showed the protein expression in red, the cytoskeleton actin was marked in green and cell nuclei were stained with TOPRO in blue. ( A1 – D1 ) p300 expression in CTRL, CurLIP, LPS-G and CurLIP/LPS-G. ( A2 – D2 ) DNMT1 expression in CTRL, CurLIP, LPS-G and CurLIP/LPS-G. ( A3 – D3 ) p300 expression in e-CTRL, e-CurLIP, e-LPS-G and CurLIP/e-LPS-G. ( A4 – D4 ) DNMT1 expression in e-CTRL, e-CurLIP, e-LPS-G and CurLIP/e-LPS-G. Scale bar = 10 µm.

Article Snippet: The primary antibodies used in the procedure were: TLR4 (1:500, Santa Cruz Biotechnology), MyD88 (1:500, Santa Cruz Biotechnology), NFkB (1:500, Santa Cruz Biotechnology), NLRP3 (3 μg/mL, Novus), caspase-1 (1:500, Santa Cruz Biotechnology), IL-1β (1 μg/mL, Termofisher), p300 (1:750, OriGene) and anti-DNMT1 (1:750, Origene); β-actin (1:750, Santa Cruz Biotechnology) was used as a normalizer to ensure loading uniformity.

Techniques: Expressing, Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Liposomal Curcumin as an Anti-Inflammatory Strategy on Lipopolysaccharide e from Porphyromonas gingivalis Treated Endothelial Committed Neural Crest Derived Stem Cells: Morphological and Molecular Mechanisms

doi: 10.3390/ijms22147534

Figure Lengend Snippet: TLR4, MyD88, NFkB, NLRP3, Caspase-1, IL-1β, DNMT1, and p300 protein quantization in hPDLSCs. ( A ) Protein levels of TLR4, MyD88, NFkB, NLRP3, Caspase-1, IL-1βDNMT1 and p300 were determined by Western blotting using specific antibodies. ( B ) Relative ratio of TLR4, MyD88, NFkB, NLRP3, Caspase-1, IL-1β, DNMT1 and p300 normalized with β-actin. *** p < 0.001; ** p < 0.01.

Article Snippet: The primary antibodies used in the procedure were: TLR4 (1:500, Santa Cruz Biotechnology), MyD88 (1:500, Santa Cruz Biotechnology), NFkB (1:500, Santa Cruz Biotechnology), NLRP3 (3 μg/mL, Novus), caspase-1 (1:500, Santa Cruz Biotechnology), IL-1β (1 μg/mL, Termofisher), p300 (1:750, OriGene) and anti-DNMT1 (1:750, Origene); β-actin (1:750, Santa Cruz Biotechnology) was used as a normalizer to ensure loading uniformity.

Techniques: Western Blot

Journal: Nature Communications

Article Title: Human iPSC-based Modeling of Pulmonary Fibrosis Reveals p300/CBP Inhibition Suppresses Alveolar Transitional Cell State

doi: 10.1038/s41467-026-68909-z

Figure Lengend Snippet: a Schematic outline of the drug screening strategy. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/h72b050 b Whole-well imaging of FD-AOs at day 17. The green areas indicate the region identified as gel using machine learning. Scale bar: 2 mm. c Comparison of manual and machine learning-based quantification of the gel area. The green area was recognized as gel. n = 20. d Dot plots showing the quantified inhibition of gel contraction in FD-AOs after treatment with 3 μg/mL BLM and 10 µM small molecules for 3 days. e Heatmap displaying the quantified inhibition of gel contraction in FD-AOs after treatment with 3 μg/mL BLM and 10 µM small molecules for 3 days. Each compound was evaluated at three different concentrations. The gray areas in the heatmap indicate doses at which cytotoxicity was observed. f Schematic outline for quantifying gel contraction in GFP + iAT2-derived FD-AOs treated with BLM. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/g85c993 g Whole-well imaging of GFP + iAT2-derived FD-AOs treated with BLM from days 11 to 14, followed by treatment with p300/CBP inhibitors from days 14 to 17. The concentration of compounds is expressed in µM. Scale bars: 2 mm. h Quantification of the matrix areas. Data are presented as mean ± SEM. One-way ANOVA followed by Tukey’s multiple comparisons test: *** p < 0.001, **** p < 0.0001. n = 3 (BLM + CBP30 1 μM, BLM + GNE781 1 μM), n = 4 (BLM + CBP30 10 μM, BLM + GNE781 10 μM), and n = 5 (DMSO, BLM) biologically independent experiments. Concentration of compounds is expressed in µM.

Article Snippet: Primary antibodies used in this study included GFP (1:500, Aves Labs, GFP-1020), SFN (1:200, Abcam, ab77187), act-p300 (1:200, biorbyt, ORB6262), EpCAM (1:200, Santa Cruz Biotechnology, sc-66020), KRT19 (1:200, Merck, MABT913), KRT17 (1:100, Abcam, ab109725), COL1A1 (1:200, Abcam, ab138492), CD54 (1:200, BioLegend, 353102), CD54 (1:200, Atlas, HPA002126), HT1-56 (1:200, Terrace Biotech, TB29AHT1-56), NaPi2b (1:200, kindly provided by Dr. Gerd Ritter (MX35)), AGER (R&D systems, AF1145), H3K27ac (1:200, Cell Signaling technology, 8173), FLAG (1:500, Cell Signaling technology, 14793), alpha smooth muscle Actin (1:100, Abcam ab5694), and Fluorescin (1:1000, Vector Laboratories, FL-1171).

Techniques: Drug discovery, Imaging, Comparison, Inhibition, Derivative Assay, Concentration Assay

Journal: Nature Communications

Article Title: Human iPSC-based Modeling of Pulmonary Fibrosis Reveals p300/CBP Inhibition Suppresses Alveolar Transitional Cell State

doi: 10.1038/s41467-026-68909-z

Figure Lengend Snippet: a Schematic outline for the isolation, sorting, and RNA-seq analysis conducted on the BLM-induced pulmonary fibrosis model of FD-AOs. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/ynmoyfa b A volcano plot derived from differential gene expression analysis of EpCAM − cells, comparing conditions with and without CBP30 (10 μM) (n = 3 biologically independent experiments). Differential expression was assessed using DESeq2 with the Wald test, and p -values were adjusted for multiple comparisons using the Benjamini–Hochberg method. Thresholds of |log₂FC| ≥ 1.5 and adjusted p -value ( p adj) ≤ 0.01 are indicated by dashed lines. EpCAM − cells were isolated from the BLM-induced pulmonary fibrosis model of FD-AOs. c Gene Ontology (GO) analysis using the top 500 downregulated genes identified by DESeq2 in EpCAM − cells comparing conditions with and without CBP30 (10 μM), ranked by adjusted p -value. GO enrichment analysis was performed using Metascape. d A volcano plot derived from differential gene expression analysis of EpCAM + cells, comparing conditions with or without CBP30 (10 μM) (n = 3 biologically independent experiments). Differential expression was assessed using DESeq2 with the Wald test, and p-values were adjusted for multiple comparisons using the Benjamini–Hochberg method. Thresholds of |log₂FC| ≥ 1.5 and p adj ≤ 0.01 are indicated by dashed lines. EpCAM + cells were isolated from the BLM FD-AOs. e Representative immunofluorescence images for Act-p300, SFN, EpCAM, and nuclei (stained with Hoechst) in FD-AOs. FD-AOs were treated with p300/CBP inhibitors (10 μM) from days 14 to 17. Scale bars: 50 μm. f Quantification of Act-p300 + cells among EpCAM + cells in the field of view (FOV). Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s test; ** p < 0.01, *** p < 0.001 ( n = 3 biologically independent experiments). g Quantification of SFN + cells in the FOV. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test; ** p < 0.01, *** p < 0.001 ( n = 3 biologically independent experiments).

Article Snippet: Primary antibodies used in this study included GFP (1:500, Aves Labs, GFP-1020), SFN (1:200, Abcam, ab77187), act-p300 (1:200, biorbyt, ORB6262), EpCAM (1:200, Santa Cruz Biotechnology, sc-66020), KRT19 (1:200, Merck, MABT913), KRT17 (1:100, Abcam, ab109725), COL1A1 (1:200, Abcam, ab138492), CD54 (1:200, BioLegend, 353102), CD54 (1:200, Atlas, HPA002126), HT1-56 (1:200, Terrace Biotech, TB29AHT1-56), NaPi2b (1:200, kindly provided by Dr. Gerd Ritter (MX35)), AGER (R&D systems, AF1145), H3K27ac (1:200, Cell Signaling technology, 8173), FLAG (1:500, Cell Signaling technology, 14793), alpha smooth muscle Actin (1:100, Abcam ab5694), and Fluorescin (1:1000, Vector Laboratories, FL-1171).

Techniques: Isolation, RNA Sequencing, Derivative Assay, Gene Expression, Quantitative Proteomics, Immunofluorescence, Staining

Journal: Nature Communications

Article Title: Human iPSC-based Modeling of Pulmonary Fibrosis Reveals p300/CBP Inhibition Suppresses Alveolar Transitional Cell State

doi: 10.1038/s41467-026-68909-z

Figure Lengend Snippet: a Schematic outline of the mouse experiments. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/b13d333 b Gene expression levels of fibrotic markers in whole lung samples under different conditions. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test, with significance levels indicated as * p < 0.05 and ** p < 0.01. n = 6 (Saline), n = 8 (BLM), and n = 8 (BLM + CBP30) independent mice. c Gene expression levels of alveolar transitional cell state markers in whole lung samples under various conditions. Data are presented as mean ± SEM. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s multiple comparisons test, with significance indicated as * p < 0.05, ** p < 0.01, and *** p < 0.001. n = 6 (Saline), n = 8 (BLM), and n = 8 (BLM + CBP30) independent mice. d Representative immunofluorescence images showing Act-p300, SFN, nuclei (Hoechst staining), and LEL in mice with BLM-induced lung injury on day 7. Scale bar: 200 μm; inset scale bar: 10 μm. e Quantification of Act-p300 + cells in the field of view (FOV). Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test, with significance levels of *** p < 0.001 and **** p < 0.0001 ( n = 3 independent mice). f Representative immunofluorescence images showing ACTA2, SFN, nuclei (Hoechst staining), and LEL in mice with BLM-induced lung injury on day 7. Scale bar: 200 μm. g, h Quantification of the ratio of ACTA2 coverage per area and the number of SFN + cells relative to total cells. Data are presented as mean ± SEM. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s multiple comparisons test, with significance levels of * p < 0.05, ** p < 0.01, and *** p < 0.001 ( n = 3 independent mice). i Working model illustrating the mode of action of p300/CBP inhibitors in pulmonary fibrosis. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/pkuir8i .

Article Snippet: Primary antibodies used in this study included GFP (1:500, Aves Labs, GFP-1020), SFN (1:200, Abcam, ab77187), act-p300 (1:200, biorbyt, ORB6262), EpCAM (1:200, Santa Cruz Biotechnology, sc-66020), KRT19 (1:200, Merck, MABT913), KRT17 (1:100, Abcam, ab109725), COL1A1 (1:200, Abcam, ab138492), CD54 (1:200, BioLegend, 353102), CD54 (1:200, Atlas, HPA002126), HT1-56 (1:200, Terrace Biotech, TB29AHT1-56), NaPi2b (1:200, kindly provided by Dr. Gerd Ritter (MX35)), AGER (R&D systems, AF1145), H3K27ac (1:200, Cell Signaling technology, 8173), FLAG (1:500, Cell Signaling technology, 14793), alpha smooth muscle Actin (1:100, Abcam ab5694), and Fluorescin (1:1000, Vector Laboratories, FL-1171).

Techniques: Gene Expression, Saline, Immunofluorescence, Staining

Journal: Nature Communications

Article Title: Human iPSC-based Modeling of Pulmonary Fibrosis Reveals p300/CBP Inhibition Suppresses Alveolar Transitional Cell State

doi: 10.1038/s41467-026-68909-z

Figure Lengend Snippet: a Gene expression of ATCS markers in the micro-patterned culture. Each well was treated with either DMSO or p300/CBP inhibitors (10 μM) from days 11 to 14. Data are presented as mean ± SEM (n = 3 biologically independent experiments). One-way ANOVA followed by Tukey’s multiple comparisons test; ** p < 0.01, *** p < 0.001, **** p < 0.0001. b Dot plots displaying the gene expression of iATCs-specific markers in representative cell populations from the micro-patterned culture, corresponding to those shown in Fig. . c Immunostaining of CD54 (ICAM1), lineage markers for ATCS (KRT19), AT1 cells (HT1-56), AT2 cells (NaPi2b), and nuclei (Hoechst) in micro-patterned cultures. Representative images from three biologically independent experiments with similar results are shown. Scale bar: 100 μm. d, e Flow cytometry analysis assessing the CD54 + cell ratio in the micro-patterned culture. Each well was treated with either DMSO or p300/CBP inhibitors (10 μM) from days 11 to 14. Data are presented as mean ± SEM (n = 3 biologically independent experiments). One-way ANOVA followed by Tukey’s multiple comparisons test; ** p < 0.01. f Schematic outline for sorting CD54 + iATCs from day 14 of the micro-patterned culture. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/v51i925 g Gene expression data of ATCS markers in the micro-patterned culture. Data are presented as mean ± SEM ( n = 3 biologically independent experiments). One-way ANOVA followed by Tukey’s multiple comparisons test; ** p < 0.01, *** p < 0.001, **** p < 0.0001. h Schematic outline of the co-culture experiment involving isolated CD54 + iATCs and NHLFs. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/nbi6c0b i Gene expression analysis of ATCS markers in isolated CD54 + iATCs. Data are presented as mean ± SEM ( n = 3 biologically independent experiments). One-way ANOVA followed by Tukey’s multiple comparisons test; ** p < 0.01. ns; not significant ( p > 0.05). j Gene expression analysis of fibroblast activation markers in NHLFs cocultured with or without iATCs. Data are presented as mean ± SEM ( n = 3 biologically independent experiments). Unpaired two-tailed Student’s t test: ∗ p < 0.05.

Article Snippet: Primary antibodies used in this study included GFP (1:500, Aves Labs, GFP-1020), SFN (1:200, Abcam, ab77187), act-p300 (1:200, biorbyt, ORB6262), EpCAM (1:200, Santa Cruz Biotechnology, sc-66020), KRT19 (1:200, Merck, MABT913), KRT17 (1:100, Abcam, ab109725), COL1A1 (1:200, Abcam, ab138492), CD54 (1:200, BioLegend, 353102), CD54 (1:200, Atlas, HPA002126), HT1-56 (1:200, Terrace Biotech, TB29AHT1-56), NaPi2b (1:200, kindly provided by Dr. Gerd Ritter (MX35)), AGER (R&D systems, AF1145), H3K27ac (1:200, Cell Signaling technology, 8173), FLAG (1:500, Cell Signaling technology, 14793), alpha smooth muscle Actin (1:100, Abcam ab5694), and Fluorescin (1:1000, Vector Laboratories, FL-1171).

Techniques: Gene Expression, Immunostaining, Flow Cytometry, Co-Culture Assay, Isolation, Activation Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Human iPSC-based Modeling of Pulmonary Fibrosis Reveals p300/CBP Inhibition Suppresses Alveolar Transitional Cell State

doi: 10.1038/s41467-026-68909-z

Figure Lengend Snippet: a Schematic outline of CUT&Tag analysis in p300/CBP inhibitors–treated iATCs. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/owocwcf b-c Representative enriched transcription factor motifs in H3K27ac peaks significantly reduced (FDR < 0.05) by CBP30 ( b ) or GNE781 ( c ) treatment. Significantly reduced peaks were identified using DiffBind, and motif enrichment analysis was performed using HOMER. d Identification of genes potentially co-regulated by AP-1 and HNF1B under p300/CBP inhibition. Genes associated with H3K27ac peaks reduced by p300/CBP inhibition were predicted using the GREAT program based on AP-1 and HNF1B motifs. The Venn diagram shows the overlap between AP-1– and HNF1B–associated genes. Among 325 shared genes, 126 genes were upregulated in iATCs compared with those of iAT2s and iAT1s (Fig. ). e Representative CUT&Tag tracks in IGV with or without p300/CBP inhibitors. Signal intensities were normalized to E. coli DNA . Red and blue bars indicate genomic regions containing AP-1 and HNF1B binding motifs identified by HOMER motif analysis. f Pathway analysis of genes co-regulated by AP-1 and HNF1B under p300/CBP inhibition. Genes predicted from AP-1 and HNF1B motifs in H3K27ac peaks reduced by p300/CBP inhibition and upregulated in iATCs (126 genes) were analyzed using IPA.

Article Snippet: Primary antibodies used in this study included GFP (1:500, Aves Labs, GFP-1020), SFN (1:200, Abcam, ab77187), act-p300 (1:200, biorbyt, ORB6262), EpCAM (1:200, Santa Cruz Biotechnology, sc-66020), KRT19 (1:200, Merck, MABT913), KRT17 (1:100, Abcam, ab109725), COL1A1 (1:200, Abcam, ab138492), CD54 (1:200, BioLegend, 353102), CD54 (1:200, Atlas, HPA002126), HT1-56 (1:200, Terrace Biotech, TB29AHT1-56), NaPi2b (1:200, kindly provided by Dr. Gerd Ritter (MX35)), AGER (R&D systems, AF1145), H3K27ac (1:200, Cell Signaling technology, 8173), FLAG (1:500, Cell Signaling technology, 14793), alpha smooth muscle Actin (1:100, Abcam ab5694), and Fluorescin (1:1000, Vector Laboratories, FL-1171).

Techniques: Inhibition, Binding Assay

Journal: Nature Communications

Article Title: Human iPSC-based Modeling of Pulmonary Fibrosis Reveals p300/CBP Inhibition Suppresses Alveolar Transitional Cell State

doi: 10.1038/s41467-026-68909-z

Figure Lengend Snippet: a Schematic outline of p300 CUT&Tag analysis in BLM-treated FD-AOs and representative transcription factor motifs enriched in p300 peaks significantly increased (adjusted p value < 0.05) upon BLM treatment. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/tzol3zv . Significantly increased peaks were identified using DiffBind. Motif enrichment was assessed using HOMER. b Representative p300 and H3K27ac CUT&Tag tracks visualized in IGV at injury-responsive epithelial genes ( CDKN1A , MMP7 , CLDN4 , and S100A2 ) in FD-AOs treated with DMSO or BLM. Signal intensities were normalized to E. coli DNA . Colored bars indicate genomic regions containing transcription factor motifs of interest. c Whole-well imaging of FD-AOs treated with BLM from days 11 to 14, followed by treatment with AP-1 inhibitors (SR11302, 10 μM; T-5224, 40 μM) from days 14 to 17. Scale bars: 2 mm. d Quantification of the matrix areas. Data are presented as mean ± SEM. One-way ANOVA followed by Tukey’s multiple comparisons test: ** p < 0.01, **** p < 0.0001 ( n = 3 biologically independent experiments). e Schematic outline of siRNA-based gene silencing in the micro-patterned culture. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/35r9mk4 f Gene expression data of ATF3 and HNF1B in the micro-patterned culture. Data are presented as mean ± SEM. n = 4 (siCont), n = 3 (siATF3), and n = 4 (siHNF1B) biologically independent experiments. One-way ANOVA followed by Tukey’s multiple comparisons test; *** p < 0.001, **** p < 0.0001. g Gene expression data of ATCS markers. Data are presented as mean ± SEM. n = 4 (siCont), n = 3 (siATF3), and n = 4 (siHNF1B) biologically independent experiments. One-way ANOVA followed by Tukey’s test; ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Primary antibodies used in this study included GFP (1:500, Aves Labs, GFP-1020), SFN (1:200, Abcam, ab77187), act-p300 (1:200, biorbyt, ORB6262), EpCAM (1:200, Santa Cruz Biotechnology, sc-66020), KRT19 (1:200, Merck, MABT913), KRT17 (1:100, Abcam, ab109725), COL1A1 (1:200, Abcam, ab138492), CD54 (1:200, BioLegend, 353102), CD54 (1:200, Atlas, HPA002126), HT1-56 (1:200, Terrace Biotech, TB29AHT1-56), NaPi2b (1:200, kindly provided by Dr. Gerd Ritter (MX35)), AGER (R&D systems, AF1145), H3K27ac (1:200, Cell Signaling technology, 8173), FLAG (1:500, Cell Signaling technology, 14793), alpha smooth muscle Actin (1:100, Abcam ab5694), and Fluorescin (1:1000, Vector Laboratories, FL-1171).

Techniques: Imaging, Gene Expression

Journal: Nature Communications

Article Title: Human iPSC-based Modeling of Pulmonary Fibrosis Reveals p300/CBP Inhibition Suppresses Alveolar Transitional Cell State

doi: 10.1038/s41467-026-68909-z

Figure Lengend Snippet: Working model of p300-mediated regulation of iATCs differentiation and epithelial–fibroblast crosstalk.

Article Snippet: Primary antibodies used in this study included GFP (1:500, Aves Labs, GFP-1020), SFN (1:200, Abcam, ab77187), act-p300 (1:200, biorbyt, ORB6262), EpCAM (1:200, Santa Cruz Biotechnology, sc-66020), KRT19 (1:200, Merck, MABT913), KRT17 (1:100, Abcam, ab109725), COL1A1 (1:200, Abcam, ab138492), CD54 (1:200, BioLegend, 353102), CD54 (1:200, Atlas, HPA002126), HT1-56 (1:200, Terrace Biotech, TB29AHT1-56), NaPi2b (1:200, kindly provided by Dr. Gerd Ritter (MX35)), AGER (R&D systems, AF1145), H3K27ac (1:200, Cell Signaling technology, 8173), FLAG (1:500, Cell Signaling technology, 14793), alpha smooth muscle Actin (1:100, Abcam ab5694), and Fluorescin (1:1000, Vector Laboratories, FL-1171).

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: ATF2 knockdown reinforces oxidative stress-induced apoptosis in TE7 cancer cells

doi: 10.1111/jcmm.12071

Figure Lengend Snippet: ATF2 regulates the expression of p21 WAF1 and c-Jun, and p-ATF2 Thr69/71 directly binds to the p21 WAF1 promoter in H 2 O 2 -treated TE7 cells (250 μM). ( A ) p-ATF2 Thr69/71 interacts with p-c-Jun Ser73 to form the AP-1 complex. In addition, p300 and CBP were found as p-ATF2 Thr69/71 interaction partners. Cells subjected to H 2 O 2 were lysed, and p-ATF2 Thr69/71 was immunoprecipitated using anti-p-ATF2 Thr69/71 antibody. Rabbit IgG was used as negative control. Precipitated lysates were immunoblotted for p-ATF2 Thr69/71 , p-c-Jun Ser73 and p300/CBP. ( B ) ATF2 knockdown causes a reduction in p-ATF2 Thr69/71 , ATF2, p21 WAF1 and c-Jun protein expression. Cells were transfected with ATF2 siRNA and transfection reagent (TFR) for 7 hrs prior to H 2 O 2 treatment. Thereafter, cells were grown for 3, 6, 12 and 24 hrs. Lysates were immunoblotted for p-ATF2 Thr69/71 , ATF2, p21 WAF1 and c-Jun. β -actin was used as loading control. Fold expression changes are given below the blots. ( C ) Schematic illustration of the p21 WAF1 promoter shows the putative ATF2-binding site (#), including the binding sequences AP-1, and CRE-BP/CREB −4772 to −4610 bp relative to the transcription start (+ 1; ATG, position 36.651.879), which was used for amplification. ( D ) p-ATF2 Thr69/71 binds to the −4772 to −4610 bp p21 WAF1 promoter region. Cells were treated with H 2 O 2 and grown for 1 and 3 hrs. Cells were lysed, and p-ATF2 Thr69/71 was immunoprecipitated using anti-p-ATF2 Thr69/71 antibody. The target region (#) in the p21 WAF1 promoter was analysed by semi-quantitative PCR. Rabbit IgG instead of anti-p-ATF2 Thr69/71 , an additional sample without antibody, and H 2 O served as negative controls. Fold PCR product accumulation is given below the gel photos.

Article Snippet: Immunodetection was carried out with anti-ATF2, -caspase 3, 8, 9, -c-Jun, -cyclin D1, -Bcl-2, -H3, -H2AX, -p-ATF2 Thr69/71 , -p-H3 Ser10 , -p-c-Jun Ser73 (Cell Signaling Technology Inc.), -Bax (DakoCytomation Inc., Carpinteria, CA, USA), - β -actin (Sigma-Aldrich, St. Louis, MO, USA), -Chk1 (Santa Cruz Biotechnology Inc.), -p300/CBP (Acris Antibodies GmbH, Herford, Germany), - γ -H2AX (p-H2AX Ser139 ; Millipore), -p-Chk1 Ser317 (Novus Biologicals Inc., Littleton, CO, USA), -p21 WAF1 (Merck, Darmstadt, Germany) and secondary antibodies (anti-mouse and anti-rabbit IgG peroxidase conjugated, Pierce, Rockford, IL, USA).

Techniques: Expressing, Immunoprecipitation, Negative Control, Knockdown, Transfection, Control, Binding Assay, Amplification, Real-time Polymerase Chain Reaction

Journal: European Journal of Histochemistry : EJH

Article Title: Porphyromonas gingivalis lipopolysaccharide stimulation in human periodontal ligament stem cells: Role of epigenetic modifications to the inflammation

doi: 10.4081/ejh.2017.2826

Figure Lengend Snippet: Immunofluorescence. Cells grown on coverslips were fixed and stained with the indicated primary and secondary antibodies and DAPI, and subjected to confocal microscopy. NF-kB expression in hPDLSCs (A1) and in hPDLSCs treated with LPS-G (A2). p300 expression in hPDLSCs (B1) and in hPDLSCs treated with LPS-G (B2). DNMT1 expression in hPDLSCs (C1) and in hPDLSCs treated with LPS-G (C2). Data are representative of three independent experiments. Magnification: 63x.

Article Snippet: The following primary monoclonal antibodies were used: antihuman NF-kB (1:250, rabbit) (OriGene Technologies, Inc., Rockville, MD, USA), anti-DNMT1 (1:250, rabbit) (OriGene), anti-p300 (1:250, rabbit) (OriGene).

Techniques: Immunofluorescence, Staining, Confocal Microscopy, Expressing

Journal: European Journal of Histochemistry : EJH

Article Title: Porphyromonas gingivalis lipopolysaccharide stimulation in human periodontal ligament stem cells: Role of epigenetic modifications to the inflammation

doi: 10.4081/ejh.2017.2826

Figure Lengend Snippet: Western blot analysis of NF-κB (A), NF-B in nuclear extract (B), DNMT1 (C) and p300 (D) expression in hPDLSCs and in hPDLSCs treated with LPS-G for 24 h. -actin was used as a housekeeping protein (E). Densitometric analysis of protein specific bands (F). **P<0.01; ***P<0.001.

Article Snippet: The following primary monoclonal antibodies were used: antihuman NF-kB (1:250, rabbit) (OriGene Technologies, Inc., Rockville, MD, USA), anti-DNMT1 (1:250, rabbit) (OriGene), anti-p300 (1:250, rabbit) (OriGene).

Techniques: Western Blot, Expressing

Journal: European Journal of Histochemistry : EJH

Article Title: Porphyromonas gingivalis lipopolysaccharide stimulation in human periodontal ligament stem cells: Role of epigenetic modifications to the inflammation

doi: 10.4081/ejh.2017.2826

Figure Lengend Snippet: Gene expression. Next generation sequencing demonstrated the modulation of genes expressed in untreated and LPS-G treated hPDLSC. P300, TNFAIP1, IL6ST and HDAC2 were expressed greater than 2-fold (Log2 fold change; Q<0.05). DNMT1 and HDAC1 genes were significantly downregulated in LPS-G stimulated hPDLSCs when compared with untreated cells (Q<0.05). Up-regulated transcripts are highlighted in red color; downregulated transcripts are highlighted in green color.

Article Snippet: The following primary monoclonal antibodies were used: antihuman NF-kB (1:250, rabbit) (OriGene Technologies, Inc., Rockville, MD, USA), anti-DNMT1 (1:250, rabbit) (OriGene), anti-p300 (1:250, rabbit) (OriGene).

Techniques: Gene Expression, Next-Generation Sequencing

Journal: The Journal of Biological Chemistry

Article Title: Biochemical impact of p300-mediated acetylation of replication protein A: Implications for DNA metabolic pathway choice

doi: 10.1016/j.jbc.2025.110250

Figure Lengend Snippet: Acetylation of RPA1 Subunit. A , IP-western blot analysis of RPA1 acetylation in wild-type HCT116, HCT116 p300 - and HCT116 p300 - cells rescued with EP300 expression. B , in vitro acetylated RPA (Ac-RPA) was subjected to SDS-PAGE analysis and stained using Coomassie brilliant blue (CBB). The same gel was subsequently analyzed by autoradiography (X-ray). C , in vitro acetylation of RPA and full-length p300 was visualized by Western blot analysis using an anti-acetyl lysine antibody. D , domains of Replication Protein A Subunit 1. Full length RPA was modified by in vitro acetylation using full-length p300 and subject to MS/MS mass spectrometry. Acetylated lysine residues on RPA1 and their positions are denoted. Competing modifications on the same lysine residue previously reported by proteomic studies are indicated, ubiquitination (square), sumoylation (triangle) mono-methylation (circle), and crotonylation (star).

Article Snippet: The EP300 cDNA plasmid in pcDNA3.1-p300 was a gift from Warner Greene (Addgene plasmid # 23252) ( ).

Techniques: Western Blot, Expressing, In Vitro, SDS Page, Staining, Autoradiography, Modification, Tandem Mass Spectroscopy, Mass Spectrometry, Residue, Ubiquitin Proteomics, Methylation

Journal: International journal of biological sciences

Article Title: Histone Acetyltransferase (HAT) P300/CBP Inhibitors Induce Synthetic Lethality in PTEN-Deficient Colorectal Cancer Cells through Destabilizing AKT.

doi: 10.7150/ijbs.42197

Figure Lengend Snippet: Figure 2. Inhibiting p300/CBP induced synthetic lethality in PTEN-/- colorectal cancer cells. (A) The structures of AA and C646. (B) Survival curves of PTEN+/+ and PTEN-/- HCT116 cells treated with C646. Cells were treated with C646 for 96 h. (C) Bar charts of cell viability in PTEN+/+/PTEN-/- cells after transfecting with p300 and CBP shRNAs for 72 h. (D) Phase-contrast and fluorescence images of PTEN+/+/PTEN-/- cells transfected with p300 or CBP shRNAs with GFP for 72 h. (E) Western blots of Ac-H4, H4, p-AKT Ser473, p-AKT Thr308, AKT1 and GAPDH in PTEN isogenic HCT116 pairs after treated with 0, 20,40, 80, 100 μM anacardic acid for 24 h. (F) Western blots of CBP, Ac-H4, p-AKT Ser473, AKT1 and GAPDH after knock-downing p300/CBP with shRNAs for 72 h in PTEN isogeneic HCT116 cell pairs. **P-values ≤ 0.01 in Student’s t-test.

Article Snippet: The shRNAs of p300 and CBP (#TG313197) were purchased from Origene (Rockville, MD).

Techniques: Fluorescence, Transfection, Western Blot