anti diabetic compound library  (TargetMol)

Journal: Journal for Immunotherapy of Cancer

Article Title: SOX2 promotes resistance of melanoma with PD-L1 high expression to T-cell-mediated cytotoxicity that can be reversed by SAHA

doi: 10.1136/jitc-2020-001037

Figure Lengend Snippet: Epigenetic compounds library screen identified SAHA decreased SOX2 level and eliminated SOX2-dependent tumor growth. (A) Schematic illustration of the screen strategy. MM200 cells were treated with 1000 IU/mL IFNγ and 5 µM epigenetic inhibitors for 24 hours, then the protein level of SOX2 was assessed by western blot assay. (B) Change of SOX2 protein level and mRNA according to Western blot and qPCR assay. ME4405 cells were treated with or without 5 µM SAHA for 24 hours. (C, D) Histogram of the percentage of cleaved caspase-3 positive cells assessed by flow cytometry. Vector and SOX2 OE ME4405 and MM200 cells were treated with or without 5 µM SAHA for 24 hours and then cocultured with T cells for 6–8 hours (Tumor: T=10:1). (E) The in vivo experimental layout. (F, G) Tumor growth curve and xenografts weight of melanoma tumors from C57BL/6 mice (control and SAHA, n=8). (H, K, L, O) Percentage of CD8+ T, NK, CD4+ T and Treg T cells in TILs for each group. (I, J, M, N) Percentage cytotoxic CD8+ T in total CD8+ T cells and cytotoxic CD4+ T in total CD4+ T cells. Tumor: T, Tumor cells: T cells. Error bars indicate SEM. One-way ANOVA test. NS, not significant; TILs, tumor infiltrating lymphocytes; *** p<0.001, ** p<0.01, * p<0.05.

Article Snippet: The epigenetic compounds library, inhibitors and cytokines are as listed: epigenetic compounds library (No. L1900, Selleck Chemicals, Houston, TX, USA), SAHA (No. S1047, Selleck Chemicals), Etidronate (No. S1857, Selleck Chemicals), PTP1B inhibitor (No. T4256, TargetMol, Boston, MA, USA), protein tyrosine phosphatase (PTP) inhibitor I (No. T7084, TargetMol), SPI 112 (No. T4340, TargetMol), murine IFNγ (No. 315-05-100, PeproTech, Rocky Hill, NJ, USA), human IFNγ (No. 300-02-1000, PeproTech), human IL2 (No. 200-02-100, PeproTech), antimouse PD-1 (CD279) (No. BE0146, BioXCell, West Lebanon, NH, USA), Rat IgG2a isotype control (No. BE0089, BioXCell).

Techniques: Western Blot, Flow Cytometry, Plasmid Preparation, In Vivo, Control

Journal: Toxicology and applied pharmacology

Article Title: Embryoid body test with morphological and molecular endpoints implicates potential developmental toxicity of trans -resveratrol

doi: 10.1016/j.taap.2018.07.006

Figure Lengend Snippet: Chemicals used for the present study

Article Snippet: trans -Resveratrol , 501-36-0 , Selleck (L2900; Anti-diabetes Compound Library) , 10 mM , 2A 2A.

Techniques: Concentration Assay, Drug discovery, Two-Dimensional Gel Electrophoresis

Journal: Toxicology and applied pharmacology

Article Title: Embryoid body test with morphological and molecular endpoints implicates potential developmental toxicity of trans -resveratrol

doi: 10.1016/j.taap.2018.07.006

Figure Lengend Snippet: Chemicals used for the present study

Article Snippet: trans -Resveratrol , 501-36-0 , Selleck (L2900; Anti-diabetes Compound Library) , 10 mM , 2A 2A.

Techniques: Concentration Assay, Drug discovery, Two-Dimensional Gel Electrophoresis

Journal: Toxicology and applied pharmacology

Article Title: Embryoid body test with morphological and molecular endpoints implicates potential developmental toxicity of trans -resveratrol

doi: 10.1016/j.taap.2018.07.006

Figure Lengend Snippet: Embryoid body morphogenesis is impacted by trans-resveratrol. (A, B) Morphometric parameters of Day 4 embryoid bodies (EBs) treated with resveratrol at different concentrations. Graphs show averages of relative area (white columns) and relative EDI (gray columns) with error bars of standard deviation. Numbers at the bottom (N) are numbers of EBs scored. No area or EDI value is available when EBs were dead (noted as “D”). Asterisks indicate adverse impacts, which are defined as reduction in average area or average EDI by more than 30% relative to controls. All adverse impacts are statistically significant (P < 0.01; two-sample t-test). (C) Representative images of EBs from one set of experiment, showing control EBs (0 μM) and those treated with different concentrations of resveratrol. Scale bar = 500 μm. (D) Impact of resveratrol on cell proliferation, evaluated by the cell viability assay (mean + standard deviation; n=3). Cells in monolayer culture were exposed to the compounds for 4 days. Asterisks indicate significant difference (P < 0.01; two-sample t-test) in mean relative light unit between control and compound treatment. (E) Morphometric parameters of Day 4 EBs treated with resveratrol-related compounds at different concentrations, presented in the same form at as described for (A, B) above.

Article Snippet: trans -Resveratrol , 501-36-0 , Selleck (L2900; Anti-diabetes Compound Library) , 10 mM , 2A 2A.

Techniques: Standard Deviation, Control, Viability Assay

Journal: Toxicology and applied pharmacology

Article Title: Embryoid body test with morphological and molecular endpoints implicates potential developmental toxicity of trans -resveratrol

doi: 10.1016/j.taap.2018.07.006

Figure Lengend Snippet: Resveratrol alters gene expression profiles in embryoid bodies. Expression levels of developmental regulator genes were determined by quantitative RT-PCR analyses. (A) Temporal expression profiles in EBs over the 4-day culture period. Horizontal axes represent days of culture whereas vertical axes represent relative expression levels in arbitrary units. Blue and red lines correspond to the relative expression levels (mean ± standard deviation; n=3) in control EBs and EBs treated with 10 μM trans-resveratrol, respectively. Asterisks indicate significant reduction or increase (P < 0.05; two-sample t-test) in mean relative expression levels by resveratrol treatment on a given day of EB culture. (B) Relative expression levels (mean ± standard deviation; n=3) in Day 2 EBs treated with trans-resveratrol. (C) Relative expression levels (mean ± standard deviation; n=3) in Day 2 EBs treated with resveratrol-related compounds at 10 μM. -: compound, tra: trans-resveratrol, cis: cis-resveratrol, 3g: trans-resveratrol 3-glucuronide, 4g: trans-resveratrol 4’-glucuronide, sul: trans-resveratrol 3-sulfate. Asterisks in (B, C) indicate significant reduction or increase (P < 0.05; two-sample t-test) in mean relative expression levels as compared to the control (0 in B, − in C).

Article Snippet: trans -Resveratrol , 501-36-0 , Selleck (L2900; Anti-diabetes Compound Library) , 10 mM , 2A 2A.

Techniques: Gene Expression, Expressing, Quantitative RT-PCR, Standard Deviation, Control

Journal: Toxicology and applied pharmacology

Article Title: Embryoid body test with morphological and molecular endpoints implicates potential developmental toxicity of trans -resveratrol

doi: 10.1016/j.taap.2018.07.006

Figure Lengend Snippet: Activation of SIRT1 is not sufficient to cause morphogenetic or molecular effects similar to resveratrol. (A) Morphometric parameters of Day 4 EBs treated with SRT1720. Graphs show averages of relative area and relative EDI ± standard deviation. The numbers of EBs scored are shown at the bottom (N). Area or EDI value is not available for 3.6 μM treatment, as EBs died or did not form (noted as “D”). (B) Percentage of hanging drops with different concentrations of SRT1720 that yielded live EBs. Numbers of hanging drops are indicated in columns. (C) Representative images of Day 4 EBs. In this set of experiment, nine out of 16 hanging drops with 3.2 μM SRT1720 (SRT) yielded live EBs, whereas all 16 control drops yielded live EBs. Scale bar= 500 μm. (D) Relative expression levels (mean ± standard deviation; n=3) of developmental regulator genes in Day 2 EBs treated with SRT1720 (SRT), compared to resveratrol (R; 10 μM) in the same sets of experiments. (E) Inhibition of SIRT1 by EX527. Purified SIRT1 protein is exposed to EX527 for 30 minutes and incubated with a luminogenic substrate to assess the SIRT1 activity. Luminescence intensity of EX527-treated SIRT1 is normalized by that of untreated SIRT1 (0 μM; set as 100%) and presented as SIRT1 activity (mean ± standard deviation; n=3). Asterisks indicate significant reduction (P < 0.01; two-samplet-test). (F) Morphometric parameters of EBs treated with EX527. Asterisks indicate adverse morphogenetic impacts. (G) Morphometric parameters of Day 4 EBs treated with EX527 with or without 10 μM resveratrol. Asterisks indicate adverse morphogenetic impacts. (H) Representative images of Day 4 EBs treated with/without resveratrol (R) and with/without EX527 (E). Scale bar= 500 μm. (I) Relative expression levels (mean ± standard deviation; n=3) of developmental regulator genes in Day 2 EBs treated with resveratrol (R; 10 μM) and with or without EX527 (E; 10 μM). Expression levels are normalized by the resveratrol treatment, which is shown as 100. Asterisks indicate significant difference (P < 0.05; two-sample t-test) in mean relative expression levels between R and R+E.

Article Snippet: trans -Resveratrol , 501-36-0 , Selleck (L2900; Anti-diabetes Compound Library) , 10 mM , 2A 2A.

Techniques: Activation Assay, Standard Deviation, Control, Expressing, Inhibition, Purification, Incubation, Activity Assay

Journal: Toxicology and applied pharmacology

Article Title: Embryoid body test with morphological and molecular endpoints implicates potential developmental toxicity of trans -resveratrol

doi: 10.1016/j.taap.2018.07.006

Figure Lengend Snippet: Estrogen receptor-modulating agents differ from resveratrol in the molecular impact. (A, B) Representative images (left) and morphometric parameters (right) of Day 4 EBs treated with diethylstilbestrol (DES; A) and with raloxifene (B). Graphs in (A, B) show averages of relative area (white columns) and relative EDI (gray columns) ± standard deviation. The numbers of EBs scored are shown at the bottom (N). Asterisks indicate adverse impacts. Scale bars = 500 μm. (C, D) Impact of DES (18 μM; C) and raloxifene (8 μM; D) on relative expression levels (mean ± standard deviation; n=3) of the developmental regulator genes over the 4-day culture. Asterisks indicate significant changes (P < 0.05; two-sample t-test) in mean relative expression levels by the estrogen receptor-modulating agent. (E) Comparisons of gene expression changes between resveratrol and the estrogen receptor-modulating agents. Downward magenta arrows indicate significant reduction compared to the control level, whereas upward green arrows indicate significant elevation. NS: No significance.

Article Snippet: trans -Resveratrol , 501-36-0 , Selleck (L2900; Anti-diabetes Compound Library) , 10 mM , 2A 2A.

Techniques: Standard Deviation, Expressing, Gene Expression, Control

Journal: Toxicology and applied pharmacology

Article Title: Embryoid body test with morphological and molecular endpoints implicates potential developmental toxicity of trans -resveratrol

doi: 10.1016/j.taap.2018.07.006

Figure Lengend Snippet: Pharmacological inhibitors of DNA replication cause morphogenetic and molecular effects similar to resveratrol. (A, B) Representative images (left) and morphometric parameters (right) of Day 4 EBs treated with aphidicolin (A) and hydroxyurea (B). Graphs in (A, B) show averages of relative area (white columns) and relative EDI (gray columns) ± standard deviation. The numbers of EBs scored are shown at the bottom (N). Asterisks indicate adverse impacts. Scale bars = 500 μm. (C, D) Impact of aphidicolin (0.4 μM; C) and hydroxyurea (40 μM; D) on gene expression patterns in EBs. Relative expression levels of the developmental regulator genes over the 4 days of culture are shown (mean ± standard deviation; n=3). Asterisks indicate significant change (P < 0.05; two-sample t-test) in mean relative expression levels by the pharmacological inhibitors of DNA replication. (E) Comparisons of gene expression changes between resveratrol, aphidicolin, and hydroxyurea. Downward magenta arrows indicate significant reduction compared to the control level, whereas upward green arrows indicate significant elevation. NS: No significance.

Article Snippet: trans -Resveratrol , 501-36-0 , Selleck (L2900; Anti-diabetes Compound Library) , 10 mM , 2A 2A.

Techniques: Standard Deviation, Gene Expression, Expressing, Control

Journal: Toxicology and applied pharmacology

Article Title: Embryoid body test with morphological and molecular endpoints implicates potential developmental toxicity of trans -resveratrol

doi: 10.1016/j.taap.2018.07.006

Figure Lengend Snippet: Resveratrol reduces the rate of DNA replication. (A) Evaluation of DNA replication rate by the EdU assay. P19C5 cells in monolayer culture were treated with vehicle control (1% DMSO), trans-resveratrol (10 μM), Aphidicolin (0.4 μM), hydroxyurea (40 μM), diethylstilbestrol (DES; 18 μM), and raloxifene (8 μM) for 24 hours, followed by incubation with EdU (100 μM) for 1 hour. Representative images of EdU labeling and nuclear staining (with Hoechst 33342) are shown. Scale bar = 50 μm. (B) Relative EdU intensity (mean ± standard deviation; n=3) to evaluate the rate of DNA replication under the compound exposures described in (A). Asterisks indicate significant difference (P < 0.01; two-sample t-test) in mean relative intensity between control and compound treatment. (C) Impact of compound exposures on cell proliferation, evaluated by the cell viability assay (mean + standard deviation; n=3). Cells in monolayer culture were exposed to the compounds for 4 days at the same concentrations as described in (A). Asterisks indicate significant difference (P < 0.01; two-sample t-test) in mean relative light unit between control and compound treatment.

Article Snippet: trans -Resveratrol , 501-36-0 , Selleck (L2900; Anti-diabetes Compound Library) , 10 mM , 2A 2A.

Techniques: EdU Assay, Control, Incubation, Labeling, Staining, Standard Deviation, Viability Assay