simplechip plus sonication chip kit  (Cell Signaling Technology Inc)

Journal: Acta biochimica et biophysica Sinica

Article Title: Integrated quantitative proteomics and phosphoproteomics analysis reveals USP46-POU4F1-HPSE signaling axis in the pathogenesis of Hirschsprung disease.

doi: 10.3724/abbs.2025064

Figure Lengend Snippet: Figure 7. USP46 regulates POU4F1 and its downstream target HPSE (A) JASPAR analysis was used to predict the POU4F1 binding sites in the HPSE promoter and identify the POU4F1 binding motif. (B) ChIP-qPCR revealed that USP46 enhances the DNA binding ability of POU4F1 by stabilizing it. *P < 0.05, ***P < 0.001, n = 3. (C) Dual-luciferase reporter assay results further confirmed that POU4F1 transcriptionally activates HPSE. *P < 0.05, n = 3. (D) Effects of USP46 overexpression or knockdown on the protein levels of POU4F1 and HPSE. (E) A qPCR assay was performed to investigate the effects of USP46 overexpression and knockdown on HPSE mRNA expression. ***P < 0.001, n = 3. (F) Enzymatic activity of HPSE measured under different conditions. ***P < 0.001, n = 3.

Article Snippet: Chromatin immunoprecipitation (ChIP-qPCR) To investigate whether POU4F1 binds to the HPSE promoter, ChIP was performed via the Simple ChIP Kit (Cell Signaling, Danvers, USA).

Techniques: Binding Assay, ChIP-qPCR, Luciferase, Reporter Assay, Over Expression, Knockdown, Expressing, Activity Assay