Fig. 3: VRK1 interacts with CHD1L and phosphorylates the CHD1L S122 site. | Cell Death & Disease

Fig. 3: VRK1 interacts with CHD1L and phosphorylates the CHD1L S122 site.

From: VRK1 promotes epithelial-mesenchymal transition in hepatocellular carcinoma mediated by SNAI1 via phosphorylating CHD1L

Fig. 3

A Schematic diagram of the screening approach of VRK1 interacting protein by immunoprecipitation combined with mass spectrometry analysis. B Mass spectral peptide count and coverage (%) of VRK1-interacting proteins. Peptide Num number of peptides used for characterization, Coverage (%) peptide coverage. C Huh7 cells were introduced with the control plasmid and transient overexpression of Flag-VRK1, then immunoprecipitated by Flag-VRK1 protein and confirmed its interacting proteins by Western blot. D, E Exogenous interactions between Flag-VRK1 and HA-CHD1L were detected by immunoprecipitation in Huh7 cells. F HA-CHD1L plasmid was transfected into Huh7 cells with silencing VRK1. The serine and threonine phosphorylation of HA-CHD1L protein immunoprecipitated by HA beads was detected by immunoblot with pan-Phospho Ser/Thr antibody. G The endogenous CHD1L protein was immunoprecipitated by protein A/G beads and the CHD1L phosphorylation level was determined. H Prediction analysis on the website (https://www.phosphosite.org) to identify CHD1L phosphorylation residue by VRK1. I The sequence conservation of CHD1L protein. J HA-CHD1L-S122A mutant was transfected into Huh7 cells with silencing VRK1. The serine and threonine phosphorylation of HA-CHD1L-S122A protein immunoprecipitated by HA beads was detected by immunoblot with pan-Phospho Ser/Thr antibody.

Back to article page