bet protein inhibitors jq1  (Tocris)

Journal: Cells

Article Title: Unraveling the Role of Bromodomain and Extra-Terminal Proteins in Human Uterine Leiomyosarcoma

doi: 10.3390/cells13171443

Figure Lengend Snippet: Treatment with JQ1 and I-BET 762 decreases uLMS cell viability and induces cell cycle arrest. ( A ) Cell viability of SK-UT-1 and UTSM cells were measured with trypan blue exclusion assay in the presence or absence of JQ1 and I-BET 762 for 48 h. ( B ) Treatment with JQ1 and I-BET 762 induces cell cycle arrest. Flow cytometric analysis was performed to measure the cell cycle phase distribution in SK-UT-1 uLMS cells in the presence or absence of JQ1 and I-BET 762. Cell cycle phases were marked as purple (G1), olive green (S), and light green (G2). Quantitative cell population analysis in response to JQ1 and I-BET 762 treatment was performed, respectively (right panel). DMSO group ( n = 4), JQ1 ( n = 4), I-BET 762 ( n = 4). ns: no significant difference; * p < 0.05; *** p < 0.001; **** p < 0.0001. ns: no significant difference. n indicates the number of biological samples for each group.

Article Snippet: BET protein inhibitors JQ1 and I-BET 762 were purchased from Tocris (Cat# 4499 and 6521, Minneapolis, MN, USA).

Techniques: Trypan Blue Exclusion Assay

Journal: Cells

Article Title: Unraveling the Role of Bromodomain and Extra-Terminal Proteins in Human Uterine Leiomyosarcoma

doi: 10.3390/cells13171443

Figure Lengend Snippet: Treatments with JQ1 and I-BET 762 sculpt the transcriptome of uLMS cells. Heat maps are presented to cluster DEGs (JQ1 vs. control) ( A ) and (I-BET 762 vs. control) ( B ), respectively. Volcano plots of gene expression profiles are presented for JQ1 vs. control ( C ) and I-BET 762 vs. control ( D ). The red and blue points represent upregulated and downregulated genes, respectively. The vertical dotted and the horizontal black lines represent the log (FC) cutoff and the logarithmic transformed adjusted p -value cutoff, respectively. ( E ) Upset diagram showing the intersection size of upregulated and downregulated genes across drug treatments. ( F ) Distribution of overlapped DEGs in response to JQ1 and I-BET 762 treatments. FC: fold-change.

Article Snippet: BET protein inhibitors JQ1 and I-BET 762 were purchased from Tocris (Cat# 4499 and 6521, Minneapolis, MN, USA).

Techniques: Control, Expressing, Transformation Assay

Journal: Cells

Article Title: Unraveling the Role of Bromodomain and Extra-Terminal Proteins in Human Uterine Leiomyosarcoma

doi: 10.3390/cells13171443

Figure Lengend Snippet: JQ1 and I-BET 762 altered cell cycle- and apoptosis-related gene expression in uLMS cells. RNA-seq analysis revealed the upregulation of CDKN1A and BIK and the downregulation of CDK6 and BCL2 , respectively, in uLMS cells upon BETis treatment. NS: no significant difference; * p < 0.05; *** p < 0.001; **** p < 0.0001.

Article Snippet: BET protein inhibitors JQ1 and I-BET 762 were purchased from Tocris (Cat# 4499 and 6521, Minneapolis, MN, USA).

Techniques: Expressing, RNA Sequencing Assay

Journal: Cells

Article Title: Unraveling the Role of Bromodomain and Extra-Terminal Proteins in Human Uterine Leiomyosarcoma

doi: 10.3390/cells13171443

Figure Lengend Snippet: JQ1 and I-BET 762 altered the Hedgehog pathway in uLMS cells. Hallmark analysis demonstrated the enrichment of the Hedgehog pathway in SK₋UT₋1 cells in response to JQ1 ( A ) and I-BET 762 ( B ) treatments. The key components of the Hedgehog pathway, including GLI1 ( C ), GLI2 ( D ), GLI3 ( E ), and SMO ( F ), are downregulated in response to JQ1 and I-BET 762 treatment. ** p < 0.01; *** p < 0.001; **** p < 0.0001. ns: no significant difference.

Article Snippet: BET protein inhibitors JQ1 and I-BET 762 were purchased from Tocris (Cat# 4499 and 6521, Minneapolis, MN, USA).

Techniques:

Journal: Cells

Article Title: Unraveling the Role of Bromodomain and Extra-Terminal Proteins in Human Uterine Leiomyosarcoma

doi: 10.3390/cells13171443

Figure Lengend Snippet: JQ1 and I-BET 762 altered the EMT pathway in uLMS cells. Hallmark analysis demonstrated the enrichment of the EMT pathway in SK₋UT₋1 cells in response to JQ1 ( A ) and I-BET 762 ( B ) treatments. The EMT inducers, including PDGFRβ ( C ), CCND2 ( D ), IL32 ( E ), and TNC ( F ), are downregulated in response to BETi treatment. *** p < 0.001; **** p < 0.0001.

Article Snippet: BET protein inhibitors JQ1 and I-BET 762 were purchased from Tocris (Cat# 4499 and 6521, Minneapolis, MN, USA).

Techniques:

Journal: Cells

Article Title: Unraveling the Role of Bromodomain and Extra-Terminal Proteins in Human Uterine Leiomyosarcoma

doi: 10.3390/cells13171443

Figure Lengend Snippet: JQ1 and I-BET 762 treatments altered DNA methylation-related genes in uLMS cells. RNA−seq revealed the downregulation of DNMT3A , DNMT3B , DNMT1 , TET1 , and TET2 in uLMS cells in response to JQ1 and I-BET 762 treatment. ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: BET protein inhibitors JQ1 and I-BET 762 were purchased from Tocris (Cat# 4499 and 6521, Minneapolis, MN, USA).

Techniques: DNA Methylation Assay, RNA Sequencing Assay

Journal: Cells

Article Title: Unraveling the Role of Bromodomain and Extra-Terminal Proteins in Human Uterine Leiomyosarcoma

doi: 10.3390/cells13171443

Figure Lengend Snippet: JQ1 and I-BET 762 altered the expression levels of histone modification-regulated genes in uLMS cells. RNA−seq revealed the altered expression of KDM1A , KDM1B , KDM4D , SETD6 , SETD9 , SETDB1 , SET , SETD9 , SIRT1 , and SIRT7 in uLMS cells in response to BETi treatments. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: BET protein inhibitors JQ1 and I-BET 762 were purchased from Tocris (Cat# 4499 and 6521, Minneapolis, MN, USA).

Techniques: Expressing, Modification, RNA Sequencing Assay

Journal: Cells

Article Title: Unraveling the Role of Bromodomain and Extra-Terminal Proteins in Human Uterine Leiomyosarcoma

doi: 10.3390/cells13171443

Figure Lengend Snippet: JQ1 and I-BET 762 altered the expression levels of m 6 A regulators in uLMS cells. RNA−seq revealed the altered expression of FTO ( A ), IGF2BP1 ( B ), and YTHDC2 ( C ) in uLMS cells in response to BETi treatment. ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: BET protein inhibitors JQ1 and I-BET 762 were purchased from Tocris (Cat# 4499 and 6521, Minneapolis, MN, USA).

Techniques: Expressing, RNA Sequencing Assay

Journal: Cells

Article Title: Unraveling the Role of Bromodomain and Extra-Terminal Proteins in Human Uterine Leiomyosarcoma

doi: 10.3390/cells13171443

Figure Lengend Snippet: Experimental model. Our experimental model shows that targeting BET proteins with JQ1 and I-BET 762 induces cell cycle arrest, modulates the Hedgehog and EMT pathways, and alters the TF network as well as interactions between target genes and epigenetic regulators in uLMS cells. This figure was created using the BioRender software online app ( BioRender.com ).

Article Snippet: BET protein inhibitors JQ1 and I-BET 762 were purchased from Tocris (Cat# 4499 and 6521, Minneapolis, MN, USA).

Techniques: Software

Journal: Biomedicines

Article Title: The Elevation and Impact of Peripheral Bile Acids in Chronic Lymphocytic Leukemia

doi: 10.3390/biomedicines13040874

Figure Lengend Snippet: Healthy donor T-cells (TC) cultured alone or co-cultured (CC) with CLL patient-derived B-CLL cells (2:1 B-CLL:T) ratio, stimulated with 10 μg/mL plate-bound anti-CD3, 5 μg/mL soluble anti-CD28, and 1.7 μM CpG-ODN 2006, and treated with the indicated BAs (50 µM), control BET inhibitor (JQ1; 5 µM) or vehicle control (VEH; DMSO) for 48–96 h (n = 6 patient samples). Following treatment, cells were evaluated by flow cytometry for immune molecule expression and function. ( A ) Percentages of co-cultured CLL cells expressing immune inhibitory molecules (48 h culture). ( B ) Top: percentage of CFSE-stained CD8 + T-cells that underwent cell division (96 h culture). Bottom: percentage of activated (CD69 + ) CD8 + T-cells (48 h culture). ( C ) Following 48 h culture, cells were stimulated with PMA/ionomycin for 6 h with Brefeldin-A added for the final 5 h. Top: the percentage of CD8 + T-cells with membrane-localized CD107a. Bottom: the percentage of polyfunctional CD8 + T-cells co-expressing IFN-γ and TNF-α. ( D , E ) Percentages of CD8 + T-cells expressing immune inhibitory receptors or transcription factors (48 h culture). ( D ) Left panels illustrate representative flow cytometry plots for the expression of PD1 ( top ) and TOX ( bottom ) in CD8 + T-cells. ( E ) PD1 lo /TIM3 − = progenitor-exhausted T-cells; PD1 hi /TIM3 + = terminally exhausted T-cells. Data are represented as mean ± SEM. Asterisks denote significant difference from CLL or TC + B-CLL (CC) VEH (one-way ANOVA with Dunnett’s correction for multiple comparisons). * p < 0.05, ** p < 0.01, *** p < 0.001. ACA: apocholic acid, TCA: taurocholic acid, CDCA: chenodeoxycholic acid, TCDCA: taurochenodeoxycholic acid, UDCA: ursodeoxycholic acid, 7KLCA: 7-ketolithocholic acid, HDCA: hyodeoxycholic acid, MDCA: murideoxycholic acid, IDCA: isodeoxycholic acid, DCA: deoxycholic acid, TLCA: taurolithocholic acid, TDCA: taurodeoxycholic acid.

Article Snippet: The BET inhibitor, JQ1 (Cayman Chemicals; Ann Arbor, MI, USA) was reconstituted in DMSO and used as a control compound known to inhibit malignant B-cell viability and downregulate the expression of immune inhibitory factors [ , ].

Techniques: Cell Culture, Derivative Assay, Control, Flow Cytometry, Expressing, Staining, Membrane