Western Blot Analysis of Epigenetic Regulators

For Western blot assays, conducted as previously described [7 (link)], antibodies against the following proteins were used: KDM4A (Novus, Littleton, CO, USA, NB110–40585), KDM4B (Bethyl Laboratories, Montgomery, TX, USA, A301–478A), KDM4C (Novus, NBP1–49600), KDM4D (Abcam, Hong Kong, China, ab198209), AR (Santa Cruz, sc-7305), histone H3 trimethyllysine 9 (H3K9me3) (Abcam, ab8898), histone H3 trimethyl lysine 4 (H3K4me3) (Abcam, ab8580), H3 (Abcam, ab1791), c-myc (Santa Cruz, sc-789), and p27^kip1 (Santa Cruz, sc-508). Protein bands were visualized by enhanced chemiluminescence (Pierce Biotechnology, Rockford, IL, USA), and protein expression was normalized against β-actin (Cell Signaling Technology, Danvers, MA, USA).

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Qiu M.T., Fan Q., Zhu Z., Kwan S.Y., Chen L., Chen J.H., Ying Z.L., Zhou Y., Gu W., Wang L.H., Cheng W.W., Zeng J., Wan X.P., Mok S.C., Wong K.K, & Bao W. (2015). KDM4B and KDM4A promote endometrial cancer progression by regulating androgen receptor, c-myc, and p27kip1. Oncotarget, 6(31), 31702-31720.

Publication 2015

KDM4A KDM4C KDM4D c-myc p27kip1 enhanced chemiluminescence β-actin

A301 Actin Antibodies C myc Chemiluminescence H3k4me3 Histone h3 Lysine Novus P27kip1 Protein Trimethyllysine Western blot assays

Corresponding Organization :

Other organizations : International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University, The University of Texas MD Anderson Cancer Center, Shanghai First Maternity and Infant Hospital, Shanghai First People's Hospital

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Variable analysis

independent variables

Antibodies against the following proteins: KDM4A, KDM4B, KDM4C, KDM4D, AR, histone H3 trimethyllysine 9 (H3K9me3), histone H3 trimethyl lysine 4 (H3K4me3), H3, c-myc, and p27kip1

dependent variables

Protein expression levels

control variables

Western blot assays conducted as previously described [7]
Protein bands visualized by enhanced chemiluminescence
Protein expression normalized against β-actin

controls

Positive control: None specified
Negative control: None specified

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