Journal: Journal of Medicinal Chemistry

Article Title: Iterative Design and Optimization of Initially Inactive Proteolysis Targeting Chimeras (PROTACs) Identify VZ185 as a Potent, Fast, and Selective von Hippel–Lindau (VHL) Based Dual Degrader Probe of BRD9 and BRD7

doi: 10.1021/acs.jmedchem.8b01413

Figure Lengend Snippet: Compound 26 induces rapid and profound depletion of BRD9 in cells. (A) Western blot analysis of BRD9, BRD7, and β-actin after treatment of HeLa cells with 1 μM compounds for 4 h before harvesting (data reported in Table ). (B) Chemical structure of 26 . (C) Quantification of BRD9 protein levels after treatment of HeLa with six different concentrations of 26 at 30 min and 4 h before harvesting. (D) Quantification of BRD9 protein levels after treatment with 1 μM 26 in HeLa at the desired time points. Intensity values, DC 50 , and half-lives were quantified as described in the .

Article Snippet: The following primary antibodies at the given concentration were used: anti β-actin (Cell Signaling Technology, 4970S, 13E5) 1:2000, anti-Brd9 (Bethyl A303-781A) 1:1000, anti-Brd7 (Bethyl A302-304A) 1:1000, VHL rabbit Ab (Cell Signaling Technology, no. 68547S) 1:1000, anti-CRBN (Novus Bio, NBP 1-9-1810) 1:1000, anti-actin hFAB rhodamine antibody, Bio-Rad, no. 12004164.

Techniques: Western Blot

Journal: Journal of Medicinal Chemistry

Article Title: Iterative Design and Optimization of Initially Inactive Proteolysis Targeting Chimeras (PROTACs) Identify VZ185 as a Potent, Fast, and Selective von Hippel–Lindau (VHL) Based Dual Degrader Probe of BRD9 and BRD7

doi: 10.1021/acs.jmedchem.8b01413

Figure Lengend Snippet: VZ185 induces strong and rapid degradation in a time- and dose-dependent manner. (A) Western blot analysis of BRD9, BRD7, and β-actin after treatment of RI-1 cells with six different concentrations of compounds 46 and 51 for 2 and 8 h before harvesting. (B) Chemical structures of 51 (VZ185) and 46 . (C) Quantification of protein levels relative to DMSO control after treatment with different concentration of VZ185 and DC 50 values. (D) Time-dependent experiment in RI-1 cells after treatment with 10 nM and 100 nM 51 at the desired time points. Intensity values and DC 50 were quantified as described in the .

Article Snippet: The following primary antibodies at the given concentration were used: anti β-actin (Cell Signaling Technology, 4970S, 13E5) 1:2000, anti-Brd9 (Bethyl A303-781A) 1:1000, anti-Brd7 (Bethyl A302-304A) 1:1000, VHL rabbit Ab (Cell Signaling Technology, no. 68547S) 1:1000, anti-CRBN (Novus Bio, NBP 1-9-1810) 1:1000, anti-actin hFAB rhodamine antibody, Bio-Rad, no. 12004164.

Techniques: Western Blot, Control, Concentration Assay