Journal: Molecules

Article Title: SAR by Space: Enriching Hit Sets from the Chemical Space

doi: 10.3390/molecules24173096

Figure Lengend Snippet: Investigational drugs—bromodomain 4 (BRD4) inhibitors.

Article Snippet: All TR-FRET assays and IC 50 measurements were performed using BRD4 (BD1+BD2) TR-FRET assay kit from BPS Bioscience ( www.bpsbioscience.com ) following the standard procedure [ ].

Techniques:

Journal: Molecules

Article Title: SAR by Space: Enriching Hit Sets from the Chemical Space

doi: 10.3390/molecules24173096

Figure Lengend Snippet: Known BRD4 inhibitors—analogs of the series identified in this work.

Article Snippet: All TR-FRET assays and IC 50 measurements were performed using BRD4 (BD1+BD2) TR-FRET assay kit from BPS Bioscience ( www.bpsbioscience.com ) following the standard procedure [ ].

Techniques:

Journal: Molecules

Article Title: SAR by Space: Enriching Hit Sets from the Chemical Space

doi: 10.3390/molecules24173096

Figure Lengend Snippet: Most active BRD4 inhibitors identified after TR-FRET (time-resolved fluorescence energy transfer) assay.

Article Snippet: All TR-FRET assays and IC 50 measurements were performed using BRD4 (BD1+BD2) TR-FRET assay kit from BPS Bioscience ( www.bpsbioscience.com ) following the standard procedure [ ].

Techniques: Fluorescence

Journal: Marine Drugs

Article Title: Inhibitors of BRD4 Protein from a Marine-Derived Fungus Alternaria sp. NH-F6

doi: 10.3390/md15030076

Figure Lengend Snippet: Inhibition rates of compounds 1 – 24 against BRD4 protein a .

Article Snippet: BRD4 (BD1) inhibitor screening assay kit was purchased from Cisbio Bioassays (Codolet, France).

Techniques: Inhibition

Journal: Epigenetics & Chromatin

Article Title: Development of novel cellular histone-binding and chromatin-displacement assays for bromodomain drug discovery

doi: 10.1186/s13072-015-0026-4

Figure Lengend Snippet: Pharmacological proof-of-concept for the in situ cell extraction methodology using reference BET bromodomain inhibitors. a Outline of the in situ cell extraction procedure. b 96-well plate view for BET bromodomain inhibitor treatment of HeLa cells expressing FLAG-tagged full-length BRD4. Cells were subjected to in situ cell extraction and two-color IF staining for TRIM24 ( green ) and histone H3 ( red ). The plate layout shows serial dilution of 3 BET inhibitors (JQ1, RVX-280 and I-BET) including wells with DMSO and secondary antibody controls. c Magnified IF images show decrease of BRD4 staining upon JQ-1 treatment (10 µM), while histone H3 ( red ) and Hoechst staining remains unchanged. d Quantification of drug dose responses based on IF image analysis. e Image analysis sequence using Harmony software. Left panel cell segmentation and gating strategy based on Hoechst ( blue ) staining eliminating cells of inappropriate morphology ( red on the second image ). Right panel the same cells with co-staining for BRD4 ( green ) and H3 ( red ). The mean intensity of the BRD4 signal was measured per nucleus and the average nucleus intensity was calculated from about 1000 gated cells (i.e., excluding the masked white cells on the fourth image)

Article Snippet: Moreover, the in situ cell extraction assay for BRD4 was exquisitely sensitive and appeared to distinguish between even small intrinsic differences in potency between JQ1 and I-BET 151 (Cell IC 50 of 132 nM and 86 nM, respectively, compared to BromoScan Kd values of 21 and 10 nM obtained in parallel at DiscoveRx for the recombinant purified proteins [ ]).

Techniques: In Situ, Extraction, Expressing, Staining, Serial Dilution, Sequencing, Software