Liver X receptor beta (LXR-β) is a member of the nuclear receptor family of transcription factors. LXR-β is encoded by the NR1H2 gene (nuclear receptor subfamily 1, group H, member 2). The liver X receptors (LXRs) were originally identified as orphan members of the nuclear receptor superfamily because their ligands were unknown. Like other receptors in the family, LXRs heterodimerize with retinoid X receptor and bind to specific response elements (LXREs) characterized by direct repeats separated by 4 nucleotides. Two genes, alpha (LXRA) and beta, are known to encode LXR proteins. Crystal structure of human liver X receptor β(LXRβ) forming heterodimer with its partner retinoid X receptor α(RXRα) on its cognate element, an AGGTCA direct repeat spaced by 4 nt shows an extended X-shaped arrangement, with DNA- and ligand-binding domains crossed. The LXRβ core binds DNA via canonical contacts and auxiliary DNA contacts that enhance affinity for the response element. Liver X receptor beta has been shown to interact with NCOA6[8] and Retinoid X receptor alpha.
背景文献
1. Zhang W et al. The LXRB-SREBP1 network regulates lipogenic homeostasis by controlling the synthesis of polyunsaturated fatty acids in goat mammary epithelial cells. J Anim Sci Biotechnol. 2022 Nov
2. Wolski H et al. Expression of ABCA1 Transporter and LXRA/LXRB Receptors in Placenta of Women with Late Onset Preeclampsia. J Clin Med. 2022 Aug
Western blot analysis of LXR beta on different lysates with Rabbit anti-LXR beta antibody (HA721403) at 1/1,000 dilution.
Lane 1: SW480 cell lysate (20 µg/Lane) Lane 2: HepG2 cell lysate (20 µg/Lane) Lane 3: PC-3M cell lysate (20 µg/Lane) Lane 4: A549 cell lysate (20 µg/Lane) Lane 5: HeLa cell lysate (20 µg/Lane) Lane 6: HT-29 cell lysate (20 µg/Lane) Lane 7: HCT116 cell lysate (20 µg/Lane) Lane 8: HEK-293 cell lysate (20 µg/Lane) Lane 9: Mouse brain tissue lysate (40 µg/Lane) Lane 10: Mouse liver tissue lysate (40 µg/Lane) Lane 11: Rat liver tissue lysate (40 µg/Lane)
Predicted band size: 51 kDa Observed band size: 51 kDa
Exposure time: 2 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 (HA721403) 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 1:100,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of C2C12 cells labeling LXR beta with Rabbit anti-LXR beta antibody (HA721403) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LXR beta antibody (HA721403) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of HCT116 cells labeling LXR beta with Rabbit anti-LXR beta antibody (HA721403) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LXR beta antibody (HA721403) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
☑ Knockdown (KD)
Western blot analysis of LXR beta on different lysates with Rabbit anti-LXR beta antibody (HA721403) at 1/1,000 dilution.
Lane 1: HEK-293-si NT cell lysate Lane 2: HEK-293-si LXR beta cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 51 kDa Observed band size: 51 kDa
Exposure time: 2 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 (HA721403) 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 1:100,000 dilution was used for 1 hour at room temperature.
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