Fig. 4: CHD1L inhibits ZKSCAN3 transcription.
From: CHD1L augments autophagy-mediated migration of hepatocellular carcinoma through targeting ZKSCAN3
A Visualization results of CHIP-seq by anti-CHD1L. Quantitative real-time-PCR (qRT–PCR) and WB were performed to determine the impact of CHD1L knockout or overexpression on mRNA (B) and protein (C) expression of ZKSCAN3 in Huh7 and QGY-7703 cells (***p < 0.001, ****p < 0.0001, Student’s t test). The Dox-induced CRISPR/Cas9 system (1 μg/ml, 72 h) was applied to knockout CHD1L in Huh7 and QGY-7703 cells, and the β-actin was used as loading control (referred to as ACTIN). D Representative images of CHD1L and ZKSCAN3 immunofluorescence staining in QGY-7703 cells transfected with GFP-tagged CHD1L. The stained cells were observed using confocal fluorescence microscopy. Scale bar represents: 10 μM. Images/immunoblots are representative of three independent experiments. Dox, Doxycycline. E, F Predicted CHD1L-binding site (ZKSCAN3 DP1, DP2 and DP3) within a region between 4741 to 4341 bp upstream of the TSS of ZKSCAN3 was evaluated by ChIP assay with anti-CHD1L IP in QGY-7703 cells. Anti-IgG IP was used as a negative control (E). ChIP-PCR was used to validate the binding of CHD1L to the identified promoter region of ZKSCAN3 in QGY-7703 cells (F). G Luciferase reporter assay in QGY-7703 cells using different fragments of ZKSCAN3 promoter after transfection with CHD1L or Vector, n = 3 biologically independent samples; data shown are mean ± SD. A two-sided Student’s t test was used to generate p values.
