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. Author manuscript; available in PMC: 2022 Jul 19.
Published in final edited form as: J Am Chem Soc. 2022 Jan 7;144(2):701–708. doi: 10.1021/jacs.1c03980

Figure 4. Characterization of FEM1B-based BRD4 degrader.

Figure 4.

(a, b) Proteasome and NEDDylation-dependence of NJH-1-106-mediated degradation of BRD4. HEK293T cells were pre-treated with DMSO vehicle, proteasome inhibitor bortezomib (BTZ) (1 μM), or NEDDylation inhibitor MLN4924 (0.2 μM) for 2 h prior to treatment with DMSO vehicle, NJH-1-106 (10 μM), or MZ1 (1 μM) for 8 h. (c) Attenuation of BRD4 degradation by EN106 and JQ1. HEK293T cells were pre-treated with DMSO vehicle, EN106 (50 μM), or JQ1 (50 μM) for 2 h prior to treatment with DMSO vehicle or NJH-1-106 (10 μM) for 8 h. (d) BRD4 degradation by NJH-1-106 in FEM1B wild-type (WT) and knockout (KO) HEK293T cells. Cells were treated with DMSO vehicle or NJH-1-106 (1 μM) for 8 h. (e) Proteomic profiling of NJH-1-106 treatment in HEK293T cells. HEK293T cells were treated with DMSO vehicle or NJH-1-106 (1 μM) for 12 h. Protein level changes in cell lysate were quantitatively assessed by TMT-based proteomic profiling. (f) Structure of FEM1B-based BCR-ABL/c-ABL degraders linking EN106 to dasatinib. (g) K562 cells were treated with DMSO vehicle or NJH-2-142 at the designated concentrations for 24 h. shown in (a-e, g) are from n=3 /group. Gels shown in (a-d, g) are representative gels from n=3 /group. Individual replicate and average ± sem values for (a-d) are shown in bar graphs. Significance is shown as *p<0.05 compared to vehicle-treated groups, and #p<0.05 compared to NJH-1-106-treated groups in (a-c) and NJH-1-106-treated WT group in (d).