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