Lysine-specific demethylase 4A is an enzyme that in humans is encoded by the KDM4A gene. This gene is a member of the Jumonji domain 2 (JMJD2) family and encodes a protein with a JmjN domain, a JmjC domain, a JD2H domain, two TUDOR domains, and two PHD-type zinc fingers. This nuclear protein belongs to the alpha-ketoglutarate-dependent hydroxylase superfamily. It functions as a trimethylation-specific demethylase, converting specific trimethylated histone on histone H3 lysine 9 and 36 residues to the dimethylated form and lysine 9 dimethylated residues to monomethyl, and as a transcriptional repressor. Alterations in this gene have been found associated with chromosomal instability that leads to cancer. In humans, the role of Kdm4a as an oncogene, or cancer associated gene, is well established. It is implicated in prostate tumors, where it is overexpressed, and stimulates cell proliferation in colon cancer cells, where it promotes formation of the tumor itself. In lung cancer cell lines, where Kdm4a is also overexpressed, it coordinates with other oncogenes (like Ras) to transform normal cells into cancerous cells by inhibiting tumor suppressor pathways such as p53. Suppression of Kdm4a in breast cancer cell lines has shown to reduce cancer cell proliferation through cell cycle arrest, and decrease tumor migration and invasion.
Background References
1. Zhang W et al. Targeting KDM4A epigenetically activates tumor-cell-intrinsic immunity by inducing DNA replication stress. Mol Cell. 2021 May
2. Sankar A et al. KDM4A regulates the maternal-to-zygotic transition by protecting broad H3K4me3 domains from H3K9me3 invasion in oocytes. Nat Cell Biol. 2020 Apr
Western blot analysis of KDM4A on different lysates with Rabbit anti-KDM4A antibody (HA721336) at 1/1,000 dilution.
Lane 1: MG-63 cell lysate (20 µg/Lane) Lane 2: Mouse cerebellum tissue lysate (30 µg/Lane) Lane 3: Mouse brain tissue lysate (30 µg/Lane) Lane 4: Rat brain tissue lysate (30 µg/Lane)
Predicted band size: 121 kDa Observed band size: 121 kDa
Exposure time: 5 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721336) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)
Western blot analysis of KDM4A on different lysates with Rabbit anti-KDM4A antibody (HA721336) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-KDM4A KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 120 kDa Observed band size: 150 kDa
Exposure time: 60 seconds; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721336) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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