Fig. 6: Targeted loss of BRD9 protein in MM leads to inhibition of cell proliferation, cell cycle progression, and colony formation and a decrease in viability.
From: BRD9 degraders as chemosensitizers in acute leukemia and multiple myeloma
A Proliferation assays: MM.1S or H929 cells treated with QA-68. 6-day assay. B Effects of QA-68 on BRD9 expression in MM.1S and H929 cells. 24-hour assay. C Effects of QA-68 on H929 cell cycle progression. 6-day assay. D Effects of QA-68 on H929 apoptosis. 6-day assay. E, upper panels Proliferation of H929 control cells and doxycycline-inducible BRD9 KD H929 cells + /-doxycycline. 4- and 6-day assays. Asterisks indicate statistical significance. E, upper left panel: t-test, 4-day: H929 control (-dox): H929 control (+dox) (P = 0.1761); H929 BRD9 KD#1 (-dox): H929 BRD9 KD#1 (+dox) (P < 0.0001). E, upper right panel: t-test, 4-day: H929 control (-dox):H929 control (+dox) (P = 0.2475); H929 BRD9 KD#2 (-dox):H929 BRD9 KD#2 (+dox) (P < 0.0001); H929 BRD9 KD#5 (-dox):H929 BRD9 KD#5 (+dox) (P < 0.0001). E, upper left panel: t-test, 6-day: H929 control (-dox):H929 control (+dox) (P = 0.9899); H929 BRD9 KD#1 (-dox):H929 BRD9 KD#1 (+dox) (P = 0.0004). E, upper right panel: t-test, 6-day: H929 control (-dox):H929 control (+dox) (P = 0.9316); H929 BRD9 KD#2 (-dox):H929 BRD9 KD#2 (+dox) (P < 0.0001); H929 BRD9 KD#5 (-dox): H929 BRD9 KD#5 (+dox) (P < 0.0003). E, lower left panel Effects of doxycycline-inducible BRD9 KD on H929 colony formation. 1000 cells/mL/well were seeded; colonies were counted 12 days postseeding. Colonies were counted twice/well. Data are presented as percent of DMSO-treated (-doxycycline) control wells. E, lower right panel BRD9 expression in H929 control cells and doxycycline-inducible BRD9 KD H929 cells + /- doxycycline. DOX, 1 µg/ml; DMSO, 0.1%. F Effects of BRD9 degrader treatment on proliferation of three primary MM patient samples versus one normal/healthy bone marrow sample, cultured in 3D tissue engineered bone marrow (3DTEBM). 5-day assay. MM bone marrow aspirates were treated with dBRD9-A (concentration range 10–1000 nM; compounds added inside 3D and outside 3D); MM survival was tested by flow cytometry (cell number normalized to counting beads, and then normalized to control). ND Newly Diagnosed, RR Relapsed/Refractory.
