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. Author manuscript; available in PMC: 2016 Jun 18.
Published in final edited form as: Chem Biol. 2015 Jun 4;22(6):755–763. doi: 10.1016/j.chembiol.2015.05.009

Figure 1. Small molecule BRD4 inhibitors lead to significant BRD4 accumulation and inefficient c-MYC suppression.

Figure 1

(A) Small molecule BRD4 inhibitors lead to significant BRD4 accumulation. Namalwa and Ramos cells were treated overnight with increasing doses of JQ1 and OTX015. Lysates were collected and subjected to immunoblot analysis with antibodies for BRD4 and actin.

(B) Small molecule BRD4 inhibitors lead to rapid BRD4 accumulation. Namalwa and Ramos cells were treated with 0.3 µM of JQ1 or OTX015 for various times as indicated, lysates were collected and analyzed by immunoblot for BRD4 and actin.

(C) Small molecule BRD4 inhibitors lead to downstream c-MYC suppression, but not efficiently. Namalwa cells were treated overnight with increasing doses of JQ1 and OTX015, lysates were collected and analyzed by immunoblot with antibodies for c-MYC and actin.

(D) Loss of c-MYC suppression shortly after BRD4 inhibitors withdrawal. (left panel) Namalwa cells were treated with JQ1 (1.0 µM) for 24 hours, followed by three washes to remove compound. Cells were re-seeded for lysates collection at various time points, c-MYC level was determined by immunoblot; (right panel) Ramos cells were treated with JQ1 (1.0 µM) or OTX015 (1.0 µM) for 24 hours, followed by compounds removal and re-seeding in fresh medium for 4 hours; lysates were subjected for immunoblot with c-MYC and actin antibodies.