Journal: iScience

Article Title: A dual SHOX2:GFP; MYH6:mCherry knockin hESC reporter line for derivation of human SAN-like cells

doi: 10.1016/j.isci.2022.104153

Figure Lengend Snippet: Generation of human SAN-like cells from hESCs (A) Schematic representation of the differentiation protocols 1–8. The protocol involves three stages (S1, S2, and S3) during the first 9 days of directed differentiation. (B–D) Quantitative PCR (B), live fluorescence images (C), and FACS plots (D) of the day 30 cells derived from the SHOX2:GFP;MYH6:mCherry H9 line using the corresponding differentiation protocols (n = at least 3 biological replicates for each condition). For quantitative PCR data, fold changes were normalized to protocol #1. p values calculated using ordinary one-way ANOVA with Fisher’s test (∗∗p < 0.01, ∗∗∗p < 0.001 ). Data are represented as mean ± SEM. The x axis in panel D is side scatter. (E) Quantitative PCR analysis for SHOX2 transcriptional expression of GFP + cells purified after sorting. n = 3 biological replicates. p value calculated by unpaired two-tailed Student’s t-test ∗∗∗p < 0.001. Data are represented as mean ± SEM, and normalized to GFP-negative population. (F) Schematic and results of the epigenetic library chemical screen. (G) Schematic of the final differentiation protocol. (H) Immunofluorescence image and FACS plot of day 30 cells derived using the protocol described in (G), Scale bar = 100 μm. AA: Activin A, B: BMP4, Ch: CHIR99021, Wi: Wnt inhibitor, RA: Retinoic Acid, Fi: FGF inhibitor, Cu: cucurbitacin, Tyr490: tyrphostin AG 490. See also Figures S1 and .

Article Snippet: Cayman Epigenetic Chemical library , Cayman , #11076.

Techniques: Real-time Polymerase Chain Reaction, Fluorescence, Derivative Assay, Expressing, Purification, Two Tailed Test, Immunofluorescence

Journal: iScience

Article Title: A dual SHOX2:GFP; MYH6:mCherry knockin hESC reporter line for derivation of human SAN-like cells

doi: 10.1016/j.isci.2022.104153

Figure Lengend Snippet:

Article Snippet: Cayman Epigenetic Chemical library , Cayman , #11076.

Techniques: Recombinant, Software

Journal: Frontiers in Pharmacology

Article Title: Epigenetic small-molecule screen for inhibition and reversal of acinar ductal metaplasia in mouse pancreatic organoids

doi: 10.3389/fphar.2024.1335246

Figure Lengend Snippet: Schematic representation of the ADM screen. Seeded organoids originating from wildtype (WT) or p48 Cre/+ (Cre) mice were treated with the Cayman small-molecule epigenetic modulator library (ESL) at final concentration of 1 µM using an automated dispensing system and left to incubate for 72 h. Following incubation, organoids were stained using the calcein AM viability dye and imaged using a high throughput imaging system at ×20 magnification to obtain high resolution images for further image analysis.

Article Snippet: We screened the Cayman’s small-molecule epigenetic modulator library (ESL) which contains 144 compounds with the goal to identify important regulators of ADM with therapeutic potential.

Techniques: Concentration Assay, Incubation, Staining, High Throughput Screening Assay, Imaging

Journal: Frontiers in Pharmacology

Article Title: Epigenetic small-molecule screen for inhibition and reversal of acinar ductal metaplasia in mouse pancreatic organoids

doi: 10.3389/fphar.2024.1335246

Figure Lengend Snippet: ADM inhibition from epigenetic compound library screen (compounds 1–68 grouped by target). The ADM inhibition assay screen was performed in duplicate using the Cayman ESL on wildtype mouse organoids. The mean percentage of viable ducts and acinar clusters (± SD) 72 h post treatment. BET = bromodomain and extraterminal domain inhibitors; DMT = DNA methyltransferase inhibitors; HDM = histone demethylase inhibitors; HMT = histone methyltransferase inhibitors; HAT = histone acetyltransferase inhibitors; HDAC = histone deacetylase inhibitors (NAD + dependent); HCl = hydrochloride; TFA salt = trifluoroacetic acid salt. p values were calculated using two-tailed Student’s t -test with unequal variances. Significance was accepted at p ≤ 0.05 only when averages of clusters are higher than the respective vehicle control. * p -value = 0.05–0.01; ** p -value = 0.01–0.001; *** p -value < 0.001. Red asterisks denote compounds that showed significantly higher cluster/duct ratios of live objects but had overall less than 50% viability of total objects (false positive). Black asterisks denote compounds that showed significantly higher cluster/duct ratios of live objects and more than 50% of total objects were viable by calcein AM staining (true positive). Single biological replicate of quadruplicate technical replicates, mean ± SD.

Article Snippet: We screened the Cayman’s small-molecule epigenetic modulator library (ESL) which contains 144 compounds with the goal to identify important regulators of ADM with therapeutic potential.

Techniques: Inhibition, Drug discovery, Histone Deacetylase Assay, Two Tailed Test, Control, Staining

Journal: Frontiers in Pharmacology

Article Title: Epigenetic small-molecule screen for inhibition and reversal of acinar ductal metaplasia in mouse pancreatic organoids

doi: 10.3389/fphar.2024.1335246

Figure Lengend Snippet: ADM inhibition from epigenetic compound library screen (compounds 69–144 grouped by target). The ADM inhibition assay screen was performed in duplicate using the Cayman ESL on wildtype mouse organoids. The mean percentage of viable ducts and acinar clusters (± SD) 72 h post treatment. HDAC = histone deacetylase inhibitors (Zn 2+ dependent); PARP = poly-ADP ribose polymerase inhibitors; HCl = hydrochloride; TFA salt = trifluoroacetic acid salt. p values were calculated using two-tailed Student’s t-test with unequal variances. Significance was accepted at p ≤ 0.05 only when averages of clusters are higher than the respective vehicle control. * p -value = 0.05–0.01; ** p -value = 0.01–0.001; *** p -value < 0.001. Red asterisks denote compounds that showed significantly higher cluster/duct ratios of live objects but had overall less than 50% viability of total objects (false positive). Black asterisks denote compounds that showed significantly higher cluster/duct ratios of live objects and more than 50% of total objects were viable by calcein AM staining (true positive). Single biological replicate of quadruplicate technical replicates, mean ± SD.

Article Snippet: We screened the Cayman’s small-molecule epigenetic modulator library (ESL) which contains 144 compounds with the goal to identify important regulators of ADM with therapeutic potential.

Techniques: Inhibition, Drug discovery, Histone Deacetylase Assay, Two Tailed Test, Control, Staining

Journal: Frontiers in Pharmacology

Article Title: Epigenetic small-molecule screen for inhibition and reversal of acinar ductal metaplasia in mouse pancreatic organoids

doi: 10.3389/fphar.2024.1335246

Figure Lengend Snippet: ADM reversal from epigenetic compound library screen (compounds 1–68 grouped by target). The ADM reversal assay screen was performed in duplicate using the Cayman ESL on Cre mouse organoids. The mean percentage of viable ducts and acinar clusters (± SD) 72 h post treatment. BET = bromodomain and extraterminal domain inhibitors; DMT = DNA methyltransferase inhibitors; HDM = histone demethylase inhibitors; HMT = histone methyltransferase inhibitors; HAT = histone acetyltransferase inhibitors; HDAC = histone deacetylase inhibitors (NAD + dependent); HCl = hydrochloride; TFA salt = trifluoroacetic acid salt. p values were calculated using two-tailed Student’s t-test with unequal variances. Significance was accepted at p ≤ 0.05 only when averages of clusters are higher than the respective vehicle control. * p -value = 0.05–0.01; ** p -value = 0.01–0.001; *** p -value < 0.001. Black asterisks denote compounds that showed significantly higher cluster/duct ratios of live objects and more than 50% of total objects were viable by calcein AM staining (true positive). Single biological replicate of quadruplicate technical replicates, mean ± SD.

Article Snippet: We screened the Cayman’s small-molecule epigenetic modulator library (ESL) which contains 144 compounds with the goal to identify important regulators of ADM with therapeutic potential.

Techniques: Drug discovery, Histone Deacetylase Assay, Two Tailed Test, Control, Staining

Journal: Frontiers in Pharmacology

Article Title: Epigenetic small-molecule screen for inhibition and reversal of acinar ductal metaplasia in mouse pancreatic organoids

doi: 10.3389/fphar.2024.1335246

Figure Lengend Snippet: ADM reversal from epigenetic compound library screen (compounds 69–144 grouped by target). The ADM reversal assay screen was performed in duplicate using the Cayman ESL on Cre mouse organoids. The mean percentage of viable ducts and acinar clusters (± SD) 72 h post treatment. HDAC = histone deacetylase inhibitors (Zn 2+ dependent); PARP = poly-ADP ribose polymerase inhibitors; HCl = hydrochloride; TFA salt = trifluoroacetic acid salt. p values were calculated using two-tailed Student’s t-test with unequal variances. Significance was accepted at p ≤ 0.05 only when averages of clusters are higher than the respective vehicle control. * p -value = 0.05–0.01; ** p -value = 0.01–0.001; *** p -value < 0.001. Red asterisks denote compounds that showed significantly higher cluster/duct ratios of live objects but had overall less than 50% viability of total objects (false positive). Black asterisks denote compounds that showed significantly higher cluster/duct ratios of live objects and more than 50% of total objects were viable by calcein AM staining (true positive). Single biological replicate of quadruplicate technical replicates, mean ± SD.

Article Snippet: We screened the Cayman’s small-molecule epigenetic modulator library (ESL) which contains 144 compounds with the goal to identify important regulators of ADM with therapeutic potential.

Techniques: Drug discovery, Histone Deacetylase Assay, Two Tailed Test, Control, Staining

Journal: NPJ Precision Oncology

Article Title: Lestaurtinib’s antineoplastic activity converges on JAK/STAT signaling to inhibit treatment naïve and therapy resistant forms ovarian cancer

doi: 10.1038/s41698-025-00947-0

Figure Lengend Snippet: a Viability of indicated cell lines following 48 h of treatment with 1 µM of the 145 compounds contained in the Cayman Chemical epigenetic drug library (#11076) relative to DMSO vehicle. N = 2. b Drug screen hit prioritization schema. Figure created with BioRender.com. c Dose response curves (two-fold dilution starting at 2.5 µM) and IC50 concentrations for lestaurtinib in a panel of therapy-sensitive and -resistant ovarian cancer cell lines following 5–10 days of treatment. N = 8. Graphs represent mean ± standard error. Abbreviations: Cis Res cisplatin-resistant, Olap R olaparib-resistant.

Article Snippet: Fig. 1 Identification of lestaurtinib as a novel inhibitor of therapy-sensitive and -resistant ovarian cancer cells. a Viability of indicated cell lines following 48 h of treatment with 1 μM of the 145 compounds contained in the Cayman Chemical epigenetic drug library (#11076) relative to DMSO vehicle.

Techniques: Drug discovery

Journal: Journal of Virology

Article Title: Histone Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid Suppresses Human Adenovirus Gene Expression and Replication

doi: 10.1128/JVI.00088-19

Figure Lengend Snippet: Pan-HDAC inhibitor SAHA suppresses HAdV late gene expression. A549 cells were infected with Ad-late/RFP (MOI of 10) and treated with vehicle, 10 μM TSA, or 10 μM SAHA. (A) Cells were fixed at 24 hpi, and the RFP fluorescence intensities were quantified using the Cellomics HCS Platform. a.u., arbitrary units. Error bars represent the standard deviations (SD) of analytical replicates (n = 16). (B) Immunoblot analysis of cell lysates collected at 24 hpi confirmed the results in panel A. Both TSA and SAHA inhibited RFP production at 24 hpi. (C and D) A549 cells (C) and MRC-5 cells (D) were infected with Ad-late/RFP and treated with 10 μM SAHA. The expression of late proteins was analyzed by immunoblotting cell lysates collected at the indicated times. SAHA significantly reduces late gene expression in both cell lines.

Article Snippet: HDAC inhibitors from the Cayman Epigenetics Screening Library that reduced RFP from Ad-late/RFP

Techniques: Gene Expression, Infection, Fluorescence, Western Blot, Expressing

Journal: Journal of Virology

Article Title: Histone Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid Suppresses Human Adenovirus Gene Expression and Replication

doi: 10.1128/JVI.00088-19

Figure Lengend Snippet: HDAC inhibitors from the Cayman Epigenetics Screening Library that reduced RFP from Ad-late/RFP

Article Snippet: HDAC inhibitors from the Cayman Epigenetics Screening Library that reduced RFP from Ad-late/RFP

Techniques:

Journal: Journal of Virology

Article Title: Histone Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid Suppresses Human Adenovirus Gene Expression and Replication

doi: 10.1128/JVI.00088-19

Figure Lengend Snippet: HDAC2 activity is necessary for efficient expression of HAdV late genes. (A) A549 cells were infected with Ad-late/RFP (MOI of 10) and treated with vehicle, SAHA, MS-275, or MC1568 (10 μM each) for 24 h. Late gene expression decreased with MS-275 but not with MC1568. (B) A549 cells were transfected with 100 nM control, HDAC1, HDAC2, or HDAC3 siRNA for 48 h. (C) Cells knocked down for each HDAC were then infected with Ad-late/RFP (MOI of 10) for 24 h to analyze fiber and RFP protein expression. (D) co-IP was performed with IgG (negative control) or E1A antibody on cells infected and drug treated as in panel A. Input was 10% of the immunoprecipitation volume. HDAC2 knockdown decreased fiber and RFP expression, and the protein was found to interact with E1A.

Article Snippet: HDAC inhibitors from the Cayman Epigenetics Screening Library that reduced RFP from Ad-late/RFP

Techniques: Activity Assay, Expressing, Infection, Gene Expression, Transfection, Control, Co-Immunoprecipitation Assay, Negative Control, Immunoprecipitation, Knockdown