Figure 7.

In cellulo assays. (A) NanoBRET assay results treating HEK293T cells for 2 h with increasing concentrations of BI7273, (B) 1–78, (C) 2–77, and (D) GSK-5959. The BRET ratio in milliBRET units (mBu) was calculated as described in Experimental Section. The IC₅₀ was calculated using the nonlinear least squares fit on GraphPad Prism 9. (E) Normalized fold increase in growth after 4 days of incubation of RWPE-1, LNCaP, 22Rv1, C4–2, PC-3, and DU145 cells in which BRD7 has been knocked down with two different constructs. Cell viability was measured with a CellTiter-Glo Luminescent Cell Viability Assay on day 0 and day 4, including 3 replicates each from 3 separate experiments. Error bars represent s.d., n = 9. (F) LNCaP and (G) PC-3 viability after 4 days of incubating the cells with compounds 1–78 and 2–77, the BRD7/9 inhibitor BI7273, and the BRPF1B inhibitor GSK-5959, including 3 replicates each from 3 separate experiments. Cell viability was measured employing a CellTiter-Glo Luminescent Cell Viability Assay. Error bars represent s.d., n = 9. (H) On-target activity assessment. 22Rv1 cells in which BRD7 has been knocked down with two different constructs were treated with 1–78, 2–77, or vehicle. Cell viability given by relative luciferase units measured with a CellTiter-Glo Luminescent Cell Viability Assay. Error bars represent s.d., n = 3. Statistical significance for panels (E)−(G) was determined using multiple t-tests with respect to the empty vector or vehicle. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, and ****p ≤ 0.0001.