Figure 1.

BRD4 Silencing Enhances Autophagic Flux

(A) Drosophila S2R⁺ cells expressing GFP-LC3 were transfected with double-stranded RNA (dsRNA) targeting control luciferase (Luc) or Fs(1)h.

(B and C) KP-4 cells transfected with control or BRD4 siRNA for 72 hr were subjected to western blot analysis (B) and stained for LC3B (C). The number of LC3 puncta normalized to cell number is shown. CON: n = 94 cells, BRD4 1: n = 97 cells, BRD4 2: n = 74 cells. Scale bars, 50 μm.

(D) Immunohistochemistry of small intestinal sections from transgenic mice harboring inducible renilla luciferase or BRD4 shRNA. Sections were stained for LC3 (upper) and BRD4 (lower). Cytoplasmic signal in BRD4 panels is due to non-specific staining. Scale bars, 50 μm.

(E) KP-4 cells transfected with BRD4 siRNA were treated with 10 μM CQ for 4 hr.

(F) KP-4 cells transfected with BRD4 siRNA were stained for WIPI2. The number of WIPI2 puncta normalized to cell number is shown. CON: n = 119 cells, BRD4 1: n = 107 cells, BRD4 2: n = 109 cells. Scale bars, 20 μm.

(G) KP-4 cells stably expressing RFP-GFP-LC3 were transfected with BRD4 siRNA. Scale bars, 50 μm.

(H) KP-4 cells were treated with 500 nM JQ1 for 9 hr in the presence or absence of CQ (10 μM, 4 hr).

(I) KP-4 cells overexpressing BRD4 were treated with 10 μM CQ for 4 hr.

(J) TY-82 cells transfected with NUT siRNA for 5 days were treated with 10 μM CQ for 8 hr. BRD4-NUT was detected using NUT antibody.

All data are shown as mean ± SD. ^∗p < 0.01. See also Figure S1.