Immunohistochemical Analysis of BET Inhibition

IHC was performed as we described previously (7 (link)). Xenograft tumor tissues were obtained from Balb/c tumor-bearing mice treated with single dose of 5 mg/kg BETd-260 or vehicle control. The following antibodies were used for IHC: BRD2 (IHC-00612), BRD4 (HC-00396), BAD (A302-384A) from Bethyl Laboratories (Shanghai, China); BRD3 (ab264420) from Abcam (Shanghai, China); cleaved PARP (Asp214) (#32563), activated caspase-3 (#9664), and Ki-67 (8D5) (9449) from Cell Signaling Technology (CST, Shanghai, China), Anti-Mcl-1 (MAB828) from R&D Systems (R&D Systems, Shanghai, China).

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Zhang H., Li G., Zhang Y., Shi J., Yan B., Tang H., Chen S., Zhang J., Wen P., Wang Z., Pang C., Li J., Guo W, & Zhang S. (2020). Targeting BET Proteins With a PROTAC Molecule Elicits Potent Anticancer Activity in HCC Cells. Frontiers in Oncology, 9, 1471.

Publication 2019

cleaved PARP (Asp214) activated caspase-3 Ki-67 (8D5)

Antibodies Balb Balb c mice Brd4 Caspase 3 Tissues Tumor Xenograft

Corresponding Organization : First Affiliated Hospital of Zhengzhou University

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Variable analysis

independent variables

Single dose of 5 mg/kg BETd-260
Vehicle control

dependent variables

Expression of BRD2, BRD3, BRD4, BAD, cleaved PARP, activated caspase-3, Mcl-1, and Ki-67 in xenograft tumor tissues

control variables

Xenograft tumor tissues obtained from Balb/c tumor-bearing mice

controls

Positive control: Not explicitly mentioned
Negative control: Vehicle control

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