rabbit anti ep300  (Cell Signaling Technology Inc)

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: