TRIM58 Interacts with ZEB1 to Suppress NSCLC Tumor Malignancy by Promoting ZEB1 Protein Degradation via UPP

Figure 4

Interaction between TRIM58 and ZEB1 was identified. (a, b) Linear regression analysis was performed to evaluate the correlation between ZEB1 protein/mRNA expression and TRIM58 protein/mRNA expression. (c) Western blot and qRT-PCR were performed to analyze ZEB1 expression in NSCLC cell lines. Each experimental cell group was treated with TRIM58-specific shRNAs, overexpression vector, or negative control vector. (d) Evaluation of TRIM58 and ZEB1 protein expression on different tumor cell groups transfected with TRIM58 specific overexpression vectors, control vectors, CQ, or MG132. (e) Protein expression quantification of the association between FLAG-tagged TRIM58 and ZEB1 protein expression level in HEK-293 T cell line. (f, g) Coimmunoprecipitation study on the protein interaction of TRIM58 with ZEB1 in NCI-H1395 cell line. (h) Immunofluorescence experiment to investigate the colocalization of TRIM58 and ZEB1 in NCI-H441 and NCI-H1395 cell lines. Overlap ratio was subsequently calculated and compared between two different cell lines.

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