Journal: OncoImmunology

Article Title: The tumor microenvironment disarms CD8+ T lymphocyte function via a miR-26a-EZH2 axis

doi: 10.1080/2162402x.2016.1245267

Figure Lengend Snippet: Fig. 3. EZH2 is a bona fide target of miR-26a. A. Schematic representation of putative

Article Snippet: In EZH2 inhibition experiments, pMel-1 CTLs were pretreated with 1-10 nM of the EZH2 inhibitor GSK-343 (Selleck Chemicals, TX, USA) or an ethanol vehicle control for 48 hours in vitro before flow cytometry analysis or intratumoral injection.

Techniques:

Journal: OncoImmunology

Article Title: The tumor microenvironment disarms CD8+ T lymphocyte function via a miR-26a-EZH2 axis

doi: 10.1080/2162402x.2016.1245267

Figure Lengend Snippet: Fig. 4. In a mouse tumor model, the miR-26a-EZH2 axis is exploited by the TME for

Article Snippet: In EZH2 inhibition experiments, pMel-1 CTLs were pretreated with 1-10 nM of the EZH2 inhibitor GSK-343 (Selleck Chemicals, TX, USA) or an ethanol vehicle control for 48 hours in vitro before flow cytometry analysis or intratumoral injection.

Techniques:

Journal: OncoImmunology

Article Title: The tumor microenvironment disarms CD8+ T lymphocyte function via a miR-26a-EZH2 axis

doi: 10.1080/2162402x.2016.1245267

Figure Lengend Snippet: Fig. 5. The human lung cancer microenvironment exploits the miR-26-EZH2 axis to

Article Snippet: In EZH2 inhibition experiments, pMel-1 CTLs were pretreated with 1-10 nM of the EZH2 inhibitor GSK-343 (Selleck Chemicals, TX, USA) or an ethanol vehicle control for 48 hours in vitro before flow cytometry analysis or intratumoral injection.

Techniques:

Journal: The Journal of Clinical Investigation

Article Title: Methyltransferase inhibitors restore SATB1 protective activity against cutaneous T cell lymphoma in mice

doi: 10.1172/JCI135711

Figure Lengend Snippet: (A) Schematic depiction of primer sites for the regulatory and control regions for ChIP-PCR analysis in B and C. (B) ChIP quantified by real-time q-PCR with anti-H3K27me3 (clone C36B11) (left) or anti-H3K9me3 (Abcam, ab8898) (right) versus the control region (approximately 14 kb from promoter) and control IgG pull-downs from HuT78 Sézary cells calculated against 2.5% input values. Regions amplified at predicted occupied region approximately 4.8 kb for H3K27me3 and approximately 5.6 kb for H3K9me3 from SATB1 promoter. Representative of 2 independent experiments. (C) ChIP quantified by real-time qPCR with anti-H3K9me3 (ab8898) and control IgG pull-downs from HuT78, Jurkat, and RAJI cells lines calculated against 2.5% input values. Regions amplified based on ChIP-seq data for H3K9me3 occupied regions near the SATB1 promoter. (D–G) HuT78 cells were treated with vehicle or increasing concentrations of the EZH2 inhibitor GSK126 for 48 hours in duplicate for MTT assay (D), SUV39H1 inhibitors chaetocin (E) and F5446 (F), or the HDAC inhibitor romidepsin (G) in duplicate, and MTT assays were performed after 72 hours. Representative of 2 independent experiments. (H–J) ChIP-PCR experiments on HuT78 cells treated with IC50 values of GSK126 (H), chaetocin (I), and romidepsin (J) pulled down with anti-H3K27me3 (clone C36B11), anti-H3K9me3 (Abcam, ab8898), or anti-H3K27ac (Abcam, ab4729), respectively. Representative of 2 independent experiments. (K) RNA was extracted from HuT78 cells identically treated for 48 hours was reversed transcribed, and SATB1 mRNA expression was quantified by q-PCR normalized to human GAPDH mRNA. Data pooled from 4 independent experiments are shown. (L) q-PCR quantification of SATB1 mRNA expression normalized to TATA-binding protein (TBP) mRNA 72 hours after treatment with the indicated doses of chaetocin, F5446, or romidepsin. Pooled from 4 independent experiments. (M) Histogram of annexin V staining on HuT78 cells treated with the IC50 value of chaetocin, F5446, or romidepsin. Two-tailed Student’s t test (B, C, and H–L): *P < 0.05; **P ≤ 0.01; ****P ≤ 0.0001.

Article Snippet: For viability analysis, HuT78 cells were seeded at 40,000 cells/well in triplicate in 96-well plates in the presence of vehicle control (ethanol or DMSO) or the EZH2 inhibitor GSK126 (Cayman Chemical, 15415), chaetocin (Abcam, ab144534), romidepsin (FK228, Depsipeptide; Selleck, S3020-5mg), F5446, and incubated at 37°C and 5% CO 2 for 48 or 72 hours.

Techniques: Control, Amplification, ChIP-sequencing, MTT Assay, Expressing, Binding Assay, Staining, Two Tailed Test

Journal: The Journal of Clinical Investigation

Article Title: Methyltransferase inhibitors restore SATB1 protective activity against cutaneous T cell lymphoma in mice

doi: 10.1172/JCI135711

Figure Lengend Snippet: (A) CD4+CD26– isolated T cells from peripheral blood apheresis of Sézary patients (n = 4) were cultured in R10 media with 100 U/mL human recombinant IL-2 and treated with increasing doses of SUV39H1/2 inhibitors chaetocin and F5446 (72 hours) as well as romidepsin (72 hours) and GSK126 (48 hours) versus the vehicle control (DMSO) prior to MTT analysis. IC50 values (nM) were calculated for each patient sample for the respective treatments. (B) Summary of IC50 values for chaetocin, F5446, romidepsin, and GSK126 for each malignant sample (n = 4). One-way ANOVA with Tukey’s multiple-comparison test: *P < 0.05; **P ≤ 0.01. (C) RNA was extracted from primary CD4+CD26– Sézary patient cells that were treated with chaetocin, F5446, romidepsin, or vehicle control (DMSO) for 24–36 hours, and SATB1 mRNA expression was quantified by q-PCR normalized to human TBP mRNA. Data pooled from 3 patient samples with 2 independent experiments shown (n = 6). Two-tailed Student’s t test: ****P ≤ 0.0001. (D) Chromatin immunoprecipitation quantified by real-time q-PCR with anti-H3K27me3 (clone C36B11) or control IgG isotype immunoprecipitation from isolated CD4+ T cells from peripheral blood of Sézary patients (n = 2) calculated against 2.5% input values. Regions amplified at the predicted occupied region (approximately 4.8 kb)of the SATB1 promoter. Representative of 2 independent experiments. Two-tailed Student’s t test: *P < 0.05; **P ≤ 0.01; ***P ≤ 0.001. (E) Similar to D except with anti-H3K9me3 (Abcam, ab8898) at the approximately 5.6-kb region versus the control region (n = 2). Representative of 2 independent experiments. Two-tailed Student’s t test: *P < 0.05; **P ≤ 0.01; ***P ≤ 0.001. (F) Primary Sézary CD4+CD26– cells (n = 2) were transduced with retrovirus containing human SATB1 and sorted for GFP+ and GFP– cells. Western blot was performed with antibodies against human p-STAT5 and STAT5 protein in cells endogenously expressing SATB1 versus cells with ectopic expression of SATB1 (n = 2). (G) Primary Sézary cells were labeled with CellTrace Violet and were serum starved for 24 hours prior to stimulation with anti-CD3/anti-CD28 beads and cultured in complete medium with 100 U/mL rhIL-2 for 5 days. Proliferation was assessed by CellTrace Violet dilution using FACS for cells ectopically (n = 2) or endogenously (n = 2) expressing SATB1. Experiment was performed on 2 replicates per patient (n = 4). Two-tailed Student’s t test: **P ≤ 0.01.

Article Snippet: For viability analysis, HuT78 cells were seeded at 40,000 cells/well in triplicate in 96-well plates in the presence of vehicle control (ethanol or DMSO) or the EZH2 inhibitor GSK126 (Cayman Chemical, 15415), chaetocin (Abcam, ab144534), romidepsin (FK228, Depsipeptide; Selleck, S3020-5mg), F5446, and incubated at 37°C and 5% CO 2 for 48 or 72 hours.

Techniques: Isolation, Cell Culture, Recombinant, Control, Comparison, Expressing, Two Tailed Test, Chromatin Immunoprecipitation, Immunoprecipitation, Amplification, Transduction, Western Blot, Labeling

Journal: Molecular cell

Article Title: Large DNA Methylation Nadirs Anchor Chromatin Loops Maintaining Hematopoietic Stem Cell Identity

doi: 10.1016/j.molcel.2020.04.018

Figure Lengend Snippet: (A). The experimental scheme. Cord blood CD34+ cells are treated with EZH2 inhibitor for 7 days in the ex vivo culture condition. The CD34+CD38- cells are isolated for in situ HiC, RNA-seq and ChIP-seq.

Article Snippet: EZH2 inhibitor-EPZ6438 , Selleckchem , S7128.

Techniques: Ex Vivo, Isolation, In Situ, RNA Sequencing, ChIP-sequencing

Journal: Molecular cell

Article Title: Large DNA Methylation Nadirs Anchor Chromatin Loops Maintaining Hematopoietic Stem Cell Identity

doi: 10.1016/j.molcel.2020.04.018

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: EZH2 inhibitor-EPZ6438 , Selleckchem , S7128.

Techniques: Recombinant, Protease Inhibitor, Transplantation Assay, DNA Methylation Assay, Purification

Journal: NPJ Regenerative Medicine

Article Title: Effective protection of photoreceptors using an inflammation-responsive hydrogel to attenuate outer retinal degeneration

doi: 10.1038/s41536-023-00342-y

Figure Lengend Snippet: a Inflammation-responsive hydrogel is formed by crosslinking of the DBCO conjugated with hyaluronic acid and azide of cathepsins-cleavable crosslinker. b The EZH2 inhibitor containing hydrogel response to cathepsins overexpressed in inflammation environment of the retina; and suppress the inflammation via disease-dependent released EZH2 inhibitor from the hydrogel.

Article Snippet: Subsequently, the prepared hydrogel, EZH2 inhibitor (Tazemetostat) of 1-2 μL was intravitreally injected into 5-week-old mice with a 33 G Hamilton needle (#NANOFIL, World Precision Instruments, Sarasota, Florida, USA).

Techniques:

Journal: NPJ Regenerative Medicine

Article Title: Effective protection of photoreceptors using an inflammation-responsive hydrogel to attenuate outer retinal degeneration

doi: 10.1038/s41536-023-00342-y

Figure Lengend Snippet: a Dextran-loaded hydrogels were incubated in PBS; control media; non-activated microglia conditioned media (NM CM); and activated microglia conditioned media (AM CM) and imaged by in vivo imaging systems (IVIS). b Relative fluorescence intensity (Region of interest; ROI) of remained dextran at each time point (mean ± SD; n = 3). c EZH2 inhibitor cumulative release profile from the hydrogel in control media; NM CM and AM CM. The supernatant containing EZH2 was measured at room temperature using UV absorbance at 256 nm (mean ± SD; n = 3). Data points within Day 1 show the short-term release profile. d In vitro release kinetics of EZH2 inhibitor from the hydrogel in PBS at 37 °C with three different cathepsins concentrations (i.e.; 0; 10; and 100 ng/mL) (mean ± SD; n = 3). Statistical analysis was performed using a one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test (*** p < 0.001).

Article Snippet: Subsequently, the prepared hydrogel, EZH2 inhibitor (Tazemetostat) of 1-2 μL was intravitreally injected into 5-week-old mice with a 33 G Hamilton needle (#NANOFIL, World Precision Instruments, Sarasota, Florida, USA).

Techniques: Incubation, Control, In Vivo Imaging, Fluorescence, In Vitro

Journal: NPJ Regenerative Medicine

Article Title: Effective protection of photoreceptors using an inflammation-responsive hydrogel to attenuate outer retinal degeneration

doi: 10.1038/s41536-023-00342-y

Figure Lengend Snippet: a mRNA level of inflammatory markers in microglia and inflammatory activated microglia (mean ± SD; n = 3). b Cathepsin L; S; and B activity in the protein level (mean ± SD; n = 3). c mRNA level of inflammatory markers in inflammatory microglia treated Gel only; Drug only; and Drug & Gel (mean ± SD; n = 3). Statistical analysis was performed using a one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test (* p < 0.1, ** p < 0.01, and *** p < 0.001). d Protein level of EZH2 associated markers in inflammatory microglia. e Schematic illustration of EZH2 inhibition associated anti-inflammatory pathway signaling.

Article Snippet: Subsequently, the prepared hydrogel, EZH2 inhibitor (Tazemetostat) of 1-2 μL was intravitreally injected into 5-week-old mice with a 33 G Hamilton needle (#NANOFIL, World Precision Instruments, Sarasota, Florida, USA).

Techniques: Activity Assay, Inhibition

Journal: Nature Communications

Article Title: Targeting p53 and histone methyltransferases restores exhausted CD8+ T cells in HCV infection

doi: 10.1038/s41467-019-14137-7

Figure Lengend Snippet: PBMC from chronically evolving (T1/early) or chronic (T2/late) HCV patients were stimulated for 40 h with HCV-NS3 peptides in the presence or absence of the EZH2 inhibitors GSK126 (GSK) and EPZ005687 (EPZ) (red dots), of the EHMT2/G9a inhibitors UNC0638 (UNC) and BIX01294 (BIX) (green dots), and of the p53 inhibitor pifithrin-alfa (p53) (blue dots). HCV-stimulated CD8+ T cells were then tested in flow cytometry for GLUT-1 levels ( a ), glucose uptake ( b ), PD-1 expression ( c ), IFN-γ ( d ), TNF-α ( e ), and IL2 ( f ) production. g – j PBMC from chronically evolving (T1/early) or chronic (T2/late) HCV patients were stimulated for 10 days with HCV-NS3 peptides in the presence or absence of the inhibitors specified in a and CD8+ T cells were then tested for IFN-γ, TNF-α, IL2, and IFN-γ plus TNF-α production as indicated. Data are presented as the ratio (fold-change) between positive CD8+ T cells detected in treated vs. untreated cultures from individual patients. Statistical analysis was performed with the Wilcoxon signed-rank test; horizontal lines represent median values. k Reduction of the repressive H3K9me2 histone mark was assessed by flow cytometry on CD8+ T cells from T2/late chronic HCV patients stimulated for 40 h or 10 days as in a . Data are presented as the ratio (fold-change) between MFI (Median fluorescence intensity) of H3K9me2 CD8+ T cells detected in treated vs. untreated cultures from individual patients; statistical analysis was performed with the Wilcoxon signed-rank test; columns and dots represent median values and single patients, respectively.

Article Snippet: The following compounds were used to restore intracellular signaling, metabolic functions and anti-viral activities: the kinase ATM inhibitor KU-55933 (tested at concentrations of 0.01–0.1 μM, Sigma), the p53 inhibitor Pifithrin-α (tested at concentrations of 10–30 μM, Sigma), the p38 inhibitor SB203580 (tested at concentrations of 0.01–0.1 μM, Sigma), the AMPK inhibitor Dorsomorphin (tested at concentrations of 0.1–1 μM, Sigma), the EZH2 inhibitor GSK126 (0.1–0.05 μΜ, Selleckchem), the EZH2 inhibitor EPZ005687 (0.1–0.05 μΜ, Selleckchem), the G9A inhibitor UNC0638 (0.1–0.5 μΜ, Selleckchem), the G9A inhibitor BIX01294 (0.5–1 μΜ, Selleckchem), and Resveratrol , (5–10 μΜ, Sigma).

Techniques: Flow Cytometry, Expressing, Fluorescence

Journal: Nature Communications

Article Title: Targeting p53 and histone methyltransferases restores exhausted CD8+ T cells in HCV infection

doi: 10.1038/s41467-019-14137-7

Figure Lengend Snippet: a Glucose uptake (measured by the incorporation of the glucose analog 2-NBDG), percentage of depolarized mitochondria (by staining with the mitochondrial membrane potential sensitive dye JC-1), PD-1 expression, proteasomal function (by ProteoStat staining) and repressive H3K9me2 mark were assessed in virus-specific, dextramer-stained CD8+ T cells from T2/late HCV patients ( n = 9) at baseline and at EOT, from T2/late resolved patients and from healthy controls after overnight anti-CD3/anti-CD28 stimulation (see Methods section for details). All data were analyzed with the Kolmogorov-Smirnov test, followed by Wilcoxon matched-pairs signed rank test (paired for chronic patients at baseline vs. EOT). Conversely, differences between multiple patient groups were evaluated with the non-parametric Kruskal-Wallis test; p -values were corrected for pairwise multiple comparisons, with the Dunn’s test. b PBMC from chronic HCV patients (T2/late; n = 6) at baseline and at EOT were stimulated for 40 h with HCV-NS3 peptides in the presence or absence of the EZH2 inhibitors GSK126 (GSK) and EPZ005687 (EPZ) (red dots), of the G9a inhibitors UNC0638 (UNC) and BIX01294 (BIX) (green dots), and of the p53 inhibitor pifithrin-alfa (p53) ( blue dots), followed by co-staining for IFN-γ, IL2, and TNFα (left). c PBMC from baseline and EOT of T2/late chronic HCV patients ( n = 11) were stimulated for 10 days with HCV-NS3 peptides as in b . Data in b and c are presented as the ratio (fold-change) between cytokine producing CD8+ T cells detected in treated vs. untreated cultures from individual patients; statistical analysis was performed with the Wilcoxon signed-rank test; horizontal lines represent median values.

Article Snippet: The following compounds were used to restore intracellular signaling, metabolic functions and anti-viral activities: the kinase ATM inhibitor KU-55933 (tested at concentrations of 0.01–0.1 μM, Sigma), the p53 inhibitor Pifithrin-α (tested at concentrations of 10–30 μM, Sigma), the p38 inhibitor SB203580 (tested at concentrations of 0.01–0.1 μM, Sigma), the AMPK inhibitor Dorsomorphin (tested at concentrations of 0.1–1 μM, Sigma), the EZH2 inhibitor GSK126 (0.1–0.05 μΜ, Selleckchem), the EZH2 inhibitor EPZ005687 (0.1–0.05 μΜ, Selleckchem), the G9A inhibitor UNC0638 (0.1–0.5 μΜ, Selleckchem), the G9A inhibitor BIX01294 (0.5–1 μΜ, Selleckchem), and Resveratrol , (5–10 μΜ, Sigma).

Techniques: Staining, Membrane, Expressing, Virus

Journal: Nature Communications

Article Title: Cathepsin-facilitated invasion of BMI1-high hepatocellular carcinoma cells drives bile duct tumor thrombi formation

doi: 10.1038/s41467-023-42930-y

Figure Lengend Snippet: BMI1 overexpression malignantly transformed liver progenitor cells into BMI1 high TICs, which are capable of initiating hepatocarcinogenesis and BDTT. Mechanistically, BMI1 collaborates with EZH2 to repress miR-218-1-3p expression by increasing H3K27me3 level, which in turn epigenetically up-regulates CTSB secretion in BMI1 high TICs to enhance their invasion into bile ducts to form tumor thrombi.

Article Snippet: For the EZH2 inhibitor treatment experiment, cells were first treated with or without 15 nM PF-06726304 (Selleck-#S8494) for 24 h prior to RT-PCR analysis.

Techniques: Over Expression, Transformation Assay, Expressing

Journal: Drug Design, Development and Therapy

Article Title:

Sevoflurane post-conditioning alleviates neonatal rat hypoxic-ischemic cerebral injury via Ezh2-regulated autophagy

doi: 10.2147/dddt.s197325

Figure Lengend Snippet: Figure 1 Long-term neurological changes after HI suffering, SPC or rapamycin injection in each group. (A) Representative cerebral morphological graphs day 34 post-HI. (B) Histological Nissil staining in CA3 region of hippocampus day 34 post-HI, scale bar =50 μm. (C) Neuronal density ratio in CA3 region of hippocampus (n=9/group), values are presented as mean±SD; ***P<0.001 compared with sham group; ###P<0.001 compared with HI group; &&&P<0.001 compared with HI + S group. Group 1: Sham (sham surgery, control), group 2: Sham + SPC (sham + S), group 3: HI, group 4: HI + SPC (HI + S), group 5: HI + SPC + rapamycin (HI + S + R), group 6: HI + rapamycin (HI + R), group 7: HI + SPC + GSK126 (HI + S + G), group 8: HI + GSK126 (HI + G). Abbreviations: SPC, sevoflurane post-conditioning; HI, hypoxic ischemia.

Article Snippet: Rats in the HI + G and HI + S + G groups received a left-side intracerebroventricular injection of the Ezh2 inhibitor GSK126 (5 μL, 50 μM; S7061, Selleck).

Techniques: Injection, Staining, Control

Journal: Drug Design, Development and Therapy

Article Title:

Sevoflurane post-conditioning alleviates neonatal rat hypoxic-ischemic cerebral injury via Ezh2-regulated autophagy

doi: 10.2147/dddt.s197325

Figure Lengend Snippet: Figure 2 Autophagy markers expression level changes 24 hrs after HI suffering, SPC or rapamycin injection in each group. Representative western blots (A) and quantification of (B) Beclin1 and (C) LC3B-II, values are presented as mean±SD (n=6/group); **P<0.01 compared with the sham group; #P<0.05, and ##P<0.01 compared with the HI group; &P<0.05, and &&P<0.01 compared with the HI + S group. (D) Representative immunofluorescent staining of LC3B (green) with NeuN (red) in hippocampus, scale bar =100 μm. Group 1: Sham (sham surgery, control), group 2: Sham + SPC (sham + S), group 3: HI, group 4: HI + SPC (HI + S), group 5: HI + SPC + rapamycin (HI + S + R), group 6: HI + rapamycin (HI + R), group 7: HI + SPC + GSK126 (HI + S + G), group 8: HI + GSK126 (HI + G). Abbreviations: SPC, sevoflurane post-conditioning; HI, hypoxic ischemia.

Article Snippet: Rats in the HI + G and HI + S + G groups received a left-side intracerebroventricular injection of the Ezh2 inhibitor GSK126 (5 μL, 50 μM; S7061, Selleck).

Techniques: Expressing, Injection, Western Blot, Staining, Control

Journal: Drug Design, Development and Therapy

Article Title:

Sevoflurane post-conditioning alleviates neonatal rat hypoxic-ischemic cerebral injury via Ezh2-regulated autophagy

doi: 10.2147/dddt.s197325

Figure Lengend Snippet: Figure 4 The Ezh2 inhibitor GSK126 reversed the SPC-conferred increase in H3K27me3, pAkt/Akt, mTOR expression, and the decrease in Pten. Representative western blots (A) and quantification of (B) Ezh2, (C) H3K27me3/H3, (D) Pten, (E) pAkt/Akt, (F) mTOR values are presented as mean±SD (n=6/group); *P<0.05, **P<0.01, ***P<0.001 compared with the sham group; #P<0.05, ##P<0.01, P<0.001 compared with the HI group; &P<0.05, and &&&P<0.001 compared with the HI + S group. Group 1: Sham (sham surgery, control), group 2: Sham + SPC (sham + S), group 3: HI, group 4: HI + SPC (HI + S), group 5: HI + SPC + rapamycin (HI + S + R), group 6: HI + rapamycin (HI + R), group 7: HI + SPC + GSK126 (HI + S + G), group 8: HI + GSK126 (HI + G). Abbreviations: SPC, sevoflurane post-conditioning; HI, hypoxic ischemia; Ezh2, Enhancer of zeste homolog 2; Pten, phosphatase and tensin homolog on chromosome 10.

Article Snippet: Rats in the HI + G and HI + S + G groups received a left-side intracerebroventricular injection of the Ezh2 inhibitor GSK126 (5 μL, 50 μM; S7061, Selleck).

Techniques: Expressing, Western Blot, Control

Journal: Drug Design, Development and Therapy

Article Title:

Sevoflurane post-conditioning alleviates neonatal rat hypoxic-ischemic cerebral injury via Ezh2-regulated autophagy

doi: 10.2147/dddt.s197325

Figure Lengend Snippet: Figure 3 The 2.5% SPC treatment for 30 mins reversed the HI-induced elevation of Pten, and the decrease in Ezh2, H3K27me3, pAkt/Akt, and mTOR expression 24 hrs post-HI. Representative western blots (A) and quantification of (B) Ezh2, (C) H3K27me3/H3, (D) Pten, (E) pAkt/Akt, and (F) mTOR values are presented as mean±SD (n=6/group); **P<0.01, and ***P<0.001 compared with the sham group; ##P<0.01, and ###P<0.001 compared with the HI group. Group 1: Sham (sham surgery, control), group 2: Sham + SPC (sham + S), group 3: HI, group 4: HI + SPC (HI + S), group 5: HI + SPC + rapamycin (HI + S + R), group 6: HI + rapamycin (HI + R), group 7: HI + SPC + GSK126 (HI + S + G), group 8: HI + GSK126 (HI + G). Abbreviations: SPC, sevoflurane post-conditioning; HI, hypoxic ischemia; Ezh2, Enhancer of zeste homolog 2; Pten, phosphatase and tensin homolog on chromosome 10.

Article Snippet: Rats in the HI + G and HI + S + G groups received a left-side intracerebroventricular injection of the Ezh2 inhibitor GSK126 (5 μL, 50 μM; S7061, Selleck).

Techniques: Expressing, Western Blot, Control

Journal: Drug Design, Development and Therapy

Article Title:

Sevoflurane post-conditioning alleviates neonatal rat hypoxic-ischemic cerebral injury via Ezh2-regulated autophagy

doi: 10.2147/dddt.s197325

Figure Lengend Snippet: Figure 5 GSK126 reverses the SPC-conferred decrease in Beclin1 and LC3B-II expression and attenuates HI-induced excessive autophagy in the CA3 hippocampal region 24 hrs post-HI. Representative western blots (A) and quantification of (B) Beclin1, (C) LC3B-II values are presented as mean±SD (n=6/group); *P<0.05, **P<0.01, and ***P<0.001 compared with the sham group; #P<0.05, and ###P<0.001 compared with the HI group; &P<0.05, &&P<0.01, and &&&P<0.001 compared with the HI + S group. One-way ANOVA with Newman-Keuls post-hoc test was used for the data analysis. (D) Representative immunofluorescent staining of LC3B. LC3B (green) showed punctate pattern in HI, HI + S + G and HI + G group, whereas sham and HI + S group presented diffused green light labeling, scale bar =100 μm. Group 1: Sham (sham surgery, control), group 2: Sham + SPC (sham + S), group 3: HI, group 4: HI + SPC (HI + S), group 5: HI + SPC + rapamycin (HI + S + R), group 6: HI + rapamycin (HI + R), group 7: HI + SPC + GSK126 (HI + S + G), group 8: HI + GSK126 (HI + G). Abbreviations: SPC, sevoflurane post-conditioning; HI, hypoxic ischemia.

Article Snippet: Rats in the HI + G and HI + S + G groups received a left-side intracerebroventricular injection of the Ezh2 inhibitor GSK126 (5 μL, 50 μM; S7061, Selleck).

Techniques: Expressing, Western Blot, Staining, Labeling, Control

Journal: Drug Design, Development and Therapy

Article Title:

Sevoflurane post-conditioning alleviates neonatal rat hypoxic-ischemic cerebral injury via Ezh2-regulated autophagy

doi: 10.2147/dddt.s197325

Figure Lengend Snippet: Figure 6 SPC improves long-term neurological outcomes partly by regulating Ezh2. (A) Representative tracing way in probe test. (B) Escape latency in place trials evaluated the acquisition of spatial information. Values are presented as mean±SD (n=10/group). (C) Time spent in each quadrant and (D) platform crossing times in probe test evaluated the memory retention ability. (E) Swimming speed in probe test and (F) suspension time performed before MWM evaluated the motor function. Numbers indicate the four quadrants of MWM, and the second quadrant was the target quadrant. Values are presented as mean±SD (n=10/group); *P<0.05, **P<0.01, ***P<0.001 compared with sham group; #P<0.05 compared with HI group; &P<0.05, &&P<0.01, &&&P<0.001 compared with HI + S group. Group 1: Sham (sham surgery, control), group 2: Sham + SPC (sham + S), group 3: HI, group 4: HI + SPC (HI + S), group 5: HI + SPC + rapamycin (HI + S + R), group 6: HI + rapamycin (HI + R), group 7: HI + SPC + GSK126 (HI + S + G), group 8: HI + GSK126 (HI + G). Abbreviations: SPC, sevoflurane post-conditioning; HI, hypoxic ischemia; MWM, Morris water maze.

Article Snippet: Rats in the HI + G and HI + S + G groups received a left-side intracerebroventricular injection of the Ezh2 inhibitor GSK126 (5 μL, 50 μM; S7061, Selleck).

Techniques: Suspension, Control

Journal: Drug Design, Development and Therapy

Article Title:

Sevoflurane post-conditioning alleviates neonatal rat hypoxic-ischemic cerebral injury via Ezh2-regulated autophagy

doi: 10.2147/dddt.s197325

Figure Lengend Snippet: Figure 7 SPC improves long-term neurological outcomes partly by regulating Ezh2. (A) Representative cerebral morphological graphs on day 34 post-HI. (B) Histological Nissil staining in CA3 region of hippocampus at day 34 after HI, scale bar =50μm. (C) Neuronal density ratio in CA3 region of hippocampus (n=9/group), values are presented as mean±SD; ***P<0.001 compared with the sham group; ###P<0.001 compared with the HI group; &&&P<0.001 compared with the HI + S group. Group 1: Sham (sham surgery, control), group 2: Sham + SPC (sham + S), group 3: HI, group 4: HI + SPC (HI + S), group 5: HI + SPC + rapamycin (HI + S + R), group 6: HI + rapamycin (HI + R), group 7: HI + SPC + GSK126 (HI + S + G), group 8: HI + GSK126 (HI + G). Abbreviations: SPC, sevoflurane post-conditioning; HI, hypoxic ischemia.

Article Snippet: Rats in the HI + G and HI + S + G groups received a left-side intracerebroventricular injection of the Ezh2 inhibitor GSK126 (5 μL, 50 μM; S7061, Selleck).

Techniques: Staining, Control

Journal: Cancers

Article Title: BET and CDK Inhibition Reveal Differences in the Proliferation Control of Sympathetic Ganglion Neuroblasts and Adrenal Chromaffin Cells

doi: 10.3390/cancers14112755

Figure Lengend Snippet: Effect of IGF, EZH2, ALK, and WNT inhibitors on the proliferation of chromaffin cells, neuroblasts, and NESTIN-expressing cells (NECs). Dose–response curves are shown for IGFR inhibitor (PPP) ( A ), EZH2 inhibitor (EPZ6438) ( B ), Alk inhibitor (Alectinib), ( C ) and Wnt inhibitor (ICG001) ( D ). Data represent the mean ± s. e. m. of at least three independent experiments.

Article Snippet: The inhibitors that were used were the BET inhibitors JQ1 (Tocris Biotechne, Wiesbaden, Germany; 4499) and GSK1324726A (iBET 726) (Selleckchem Biozol, Eching, Germany), the CDK-7 inhibitors THZ1 (Medchem Express Biotrend, Köln, Germany) and YKL-5-125 (Selleckchem), the CDK12/13 inhibitor THZ 531 (Selleckchem), the IGF1-R inhibitor picropodophyllin (PPP) (Tocris 2956), the EZH2 inhibitor EPZ 6438 (Axon Medchem, Groningen, NL; 2227), the WNT inhibitor ICG001 (Axon Medchem 1766), and the ALK inhibitor Alectinib (Selleckchem S276).

Techniques: Expressing