Additional file 5 of An SETD1A/Wnt/β-catenin feedback loop promotes NSCLC development
figureposted on 14.10.2021, 03:24 authored by Rui Wang, Jian Liu, Kai Li, Ganghua Yang, Sisi Chen, Jie Wu, Xinming Xie, Hong Ren, Yamei Pang
Additional file 5: Figure S3. The effects of SETD1A on the Wnt/β-catenin pathway. A, Wnt/β-catenin pathway activity in A549 cells was detected by TOP/FOP flash reporter assay following SETD1A knockdown and overexpression. B, the nuclear β-catenin levels were analyzed by immunofluorescence assay following transfection with the empty vector and SETD1A plasmid. C, CTNNB1 transcript levels in NSCLC cells were detected by qRT-PCR following SETD1A overexpression. D, Cytoplasmic β-catenin levels was detected by western blotting following SETD1A knockdown and overexpression. E, The subcellular localization of SETD1A protein was predicted by the BUSCA webserver. F, The interaction between SETD1A and β-catenin in BEAS2B cells was analyzed by co-immunoprecipitation. G, β-catenin stability was analyzed by CHX chase assay following transfection with the empty vector and SETD1A plasmid. H, Immunofluorescence analysis using normal IgG in PC9 cells is shown. I, β-catenin expression was attenuated by MG132 treatment in SETD1A knockdown cells. J, PC9 cell lysates were immunoprecipitated using a β-catenin antibody and subjected to western blot analysis with the corresponding antibodies as indicated after Wnt3a treatment for 12 h. K, S675 phosphorylation of β-catenin was determined by western blotting following SETD1A overexpression without Wnt3a stimulation. Data are shown as means ± SD. *P < 0.05, **P < 0.01.
Young Scientists Fund National Natural Science Foundation of China Scientific and Technological Development Research Project Foundation of Shaanxi Province