USP2 Inhibits Lung Cancer Pathogenesis by Reducing ARID2 Protein Degradation via Ubiquitination

Figure 1

USP2 is significantly suppressed and interacts with ARID2 in lung cancer cells; H1299 cells were transfected with Sh USP2#1, Sh USP2#2, Sh USP2#3, or negative control vector. A549 cells were transfected with USP2 overexpression vector or negative control vector. (a) Coomassie blue staining on H1299 cell line protein samples immunoprecipitated with IgG and anti-USP2 antibody. (b) Validation of USP2-ARID2 interaction by immunoprecipitation method in H1299 and A549 cell lines. (c) Immunofluorescence colocalization method to detect USP2-ARID2 localization in H1299 and A549 cells. (d) Statistical analysis on the colocalization ratio of USP2 and ARID2 using overlap and Pearson’s ratio parameters, respectively. (e, f) Expression profile of USP2 protein and mRNA in multiple human lung cancer cell lines (NCI-H1975, NCI-H292, H1299, NCI-H1395, and A549) and normal human bronchial epithelial cell line (HBE) by WB tests ( indicates compared with group HBE; indicates compared with group HBE). (g, h) Design and validation of USP2-specific shRNAs and USP2-specific overexpression vectors’ effects on USP2 mRNA expression in H1299 and A549 cells by PCR methods ( indicates compared with group control; indicates compared with group control or vector).

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