Fig. 5

DYRK1A activates TGF-β/SMAD signalling by interacting with TSC1. a A diagram was generated using the online tool Venny (http://www.bioinformatics.com.cn/static/others/jvenn/example.html). b An immunoprecipitation assay was carried out to assess the interaction between DYRK1A and TSC1. c and d The results were acquired from the GEPIA database (gepia.cancer-pku.cn). Gene: DYRK1A; The Cancer Genome Atlas Tumour: Liver HCC. e An immunoprecipitation assay was performed to evaluate the interaction among DYRK1A, TSC1, and SMAD2/3. f DYRK1A was knocked down in HCC cells by transfection with the corresponding siRNA for 48 h. Total protein was extracted from HCC cells and analysed by western blotting. g Following treatment with harmine for 24 h, total protein was extracted from HCC cells and analysed. h HCC cells were incubated with the DYRK1A overexpression plasmid or empty vector for 48 h, and western blot analysis was performed. i HCC cells were serum-starved overnight and were then incubated with 10 μM harmine and/or 10 ng/mL TGF-β for 6 h. Subsequently, protein levels were measured. j HCC cells were incubated with DYRK1A siRNA or control siRNA for 24 h. Subsequently, the cells were first serum-starved overnight and then incubated with DMSO or 10 ng/mL TGF-β for 6 h. Western blot analysis was then performed