rarβ antagonist le135  (Tocris)

Journal: EMBO Reports

Article Title: Rarγ-Foxa1 signaling promotes luminal identity in prostate progenitors and is disrupted in prostate cancer

doi: 10.1038/s44319-024-00335-y

Figure Lengend Snippet: Reagents and tools table

Article Snippet: RARβ antagonist (LE135) , Cayman , 14415.1.

Techniques: Recombinant, Sequencing, Software

Journal: Biomedicines

Article Title: Histone H3 Acetylation Is Involved in Retinoid Acid-Induced Neural Differentiation through Increasing Mitochondrial Function

doi: 10.3390/biomedicines11123251

Figure Lengend Snippet: RA promotes histone acetylation through RA signaling during neural differentiation of SH-SY5Y cells. ( a – c ) RT-qPCR analysis of relative mRNA expression of RARα , RARβ , and RARγ in SH-SY5Y cells after RA treatment for 3 and 7 days. n = 3; ** p < 0.01; one-way ANOVA. ( d ) Co-immunoprecipitation was performed in SH-SY5Y cells cultured in maintenance medium or stimulated with RA for 3 and 7 days. Immunoprecipitation was conducted using anti-RARα followed by Western blot analysis of PCAF, CBP, and P300. ( e – g ) Western blot analysis of H3K14ac and H3ac in SH-SY5Y cells treated with DMSO or RA or pretreated with BMS195614/LE135 for 24 h followed by RA for 3 days. n = 3; * p < 0.05; one-way ANOVA.

Article Snippet: To identify the roles of key enzymes and receptors during neural differentiation, the HAT activity inhibitor C646 (Sigma, St. Louis, MO, USA), RARα antagonist BMS195614, and RARβ antagonist LE135 (Glpbio, Montclair, CA, USA) were supplemented during the RA treatment.

Techniques: Quantitative RT-PCR, Expressing, Immunoprecipitation, Cell Culture, Western Blot

Journal: Biomedicines

Article Title: Histone H3 Acetylation Is Involved in Retinoid Acid-Induced Neural Differentiation through Increasing Mitochondrial Function

doi: 10.3390/biomedicines11123251

Figure Lengend Snippet: Inhibition of RA signaling downregulates mitochondrial function and blocks neural differentiation. SH-SY5Y cells were treated with DMSO or RA or pretreated with BMS195614/LE135 for 24 h followed by RA for 3 days. ( a – d ) RT-qPCR analysis of relative mRNA expression of mitochondrial function-related genes ( NDUFS3 , ANT1 , ANT2 , ANT3 ). n = 3; * p < 0.05, ** p < 0.01; one-way ANOVA. ( e ) Changes in morphology and expression of neuronal differentiation markers Tuj1 (green) and MAP2 (red). Scale bar, 50 µm. Images in the black box show cells at a higher magnification. Red or white arrows show representative changes in morphology. ( f – h ) Differentiation rates and mean fluorescence intensity of Tuj1 and MAP2. n = 3; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; one-way ANOVA. ( i , j ) RT-qPCR analysis of relative mRNA expression of the neural differentiation markers Tuj1 and MAP2 . n = 3; * p < 0.05, ** p < 0.01; one-way ANOVA.

Article Snippet: To identify the roles of key enzymes and receptors during neural differentiation, the HAT activity inhibitor C646 (Sigma, St. Louis, MO, USA), RARα antagonist BMS195614, and RARβ antagonist LE135 (Glpbio, Montclair, CA, USA) were supplemented during the RA treatment.

Techniques: Inhibition, Quantitative RT-PCR, Expressing, Fluorescence

Journal: Experimental dermatology

Article Title: RARα/RXR Synergism Potentiates Retinoid Responsiveness in Cutaneous T Cell Lymphoma Cell Lines

doi: 10.1111/exd.13348

Figure Lengend Snippet: RARα activation prompts β7 expression and function in CTCL. (a) MJ or HuT78 cells were treated with DMSO or 2×EC50 of well-established RAR isotype agonists for 24, 48 or 72 hrs. Surface β7 integrin expression was determined through flow cytometry and displayed as mean fluorescence intensity (MFI). (b) MJ or HuT78 cells were cultured in the presence of DMSO, 100 nM ATRA, 2×EC50 RARα, β, or γ agonists for the indicated time. Static cell adhesion assays were assessed on 0.75 μg/ml of the β7 specific ligand MAdCAM-1. (c) MJ cells were cultured with the indicated RAR isotype antagonists at 500 nM for 24 hrs. Cells were further subcultured for an additional 24 hrs in the presence of 200 nM ATRA. Static cell adhesion assays were conducted as described before. Ordinate represents data that have been normalized to adhesion levels obtained with ATRA in the absence of antagonist. (d) Adhesion assays were repeated as previously described in (B) with the non-CTCL cell line, Jurkat. (e) Whole cell lysates (30 μg/lane) of CTCL or non-CTCL (Jurkat) cell lines were examined for the presence and relative abundance of the various RAR receptor isotypes.

Article Snippet: RARα agonist AM580, RARβ agonist CD2314, and RARγ agonist BMS961 were obtained from TOCRIS bioscience through Sigma, as were the selective receptor antagonists for RARα (ER5089), RARβ/γ (CD2665), and RARβ (LE135).

Techniques: Activation Assay, Expressing, Flow Cytometry, Fluorescence, Cell Culture

Journal: Experimental dermatology

Article Title: RARα/RXR Synergism Potentiates Retinoid Responsiveness in Cutaneous T Cell Lymphoma Cell Lines

doi: 10.1111/exd.13348

Figure Lengend Snippet: RARα/RXR coactivation induces CTCL cell adhesion in a synergistic manner. (a) MJ or HuT78 cells were cultured in the presence of RAR isotype-specific agonists (1×EC50) alone or in combination with 5 nM Bexarotene for 24 hrs. Cells were then assessed for adhesion to 0.75 μg/ml of MAdCAM-1. (b) CTCL cell line SeAx or MyLa cells were cultured with DMSO, 2×EC50 RARα agonist, 100 nM Bexarotene, or a combination of agonists for 72 hrs. The extent of adhesion was determined on the integrin β7-ligand MAdCAM-1. (c) HuT78 cells were cultured in the presence of an RAR isotype-specific agonist (1×EC50) alone or in combination with 100 nM of the pan-RXR activator SR11237 for 48 hrs. Cell adhesion was then determined as previously described. (d) Real-time quantitative PCR was conducted with primers that amplify regions encoding the human integrin α4 or β7 subunits. Templates were derived from MJ cells cultured with vehicle or RARα/RXR agonists. The ordinate reflects normalized data obtained by dividing values for RARα/RXR agonists with those from vehicle treated samples.

Article Snippet: RARα agonist AM580, RARβ agonist CD2314, and RARγ agonist BMS961 were obtained from TOCRIS bioscience through Sigma, as were the selective receptor antagonists for RARα (ER5089), RARβ/γ (CD2665), and RARβ (LE135).

Techniques: Cell Culture, Real-time Polymerase Chain Reaction, Derivative Assay