Research Paper Volume 15, Issue 19 pp 10593—10606
UCHL1-PKM2 axis dysregulation is associated with promoted proliferation and invasiveness of urothelial bladder cancer cells
Figure 4. UCHL1 interacts with PKM2 and upregulates the PKM2 protein level. (A) Coomassie blue staining experiment on anti-UCHL1-immunoprecipitated cell protein samples, and Co-IP-MS assay was performed on samples immunoprecipitated by anti-UCHL1 antibody to investigate potential UCHL1 interacting protein targets. (B) Co-IP assay to confirm the protein-protein interaction of UCHL1 and PKM2 in T24 and J82 cell line models. (C) Immunofluorescence co-localization assay to confirm the protein interaction of UCHL1 and PKM2 in T24 and J82 cell line models. (D) Evaluation of UCHL1 regulatory role on PKM2 by exogenously transfection of His-tagged UCHL1 and His-tagged loss-of-function UCHL1 C90S mutated vector into HEK-293T cells. (E, F) Explore the impact of UCHL1 silencing or overexpression on PKM2 expression in UBC cancer cell line models by transfection of UCHL1-specific shRNAs or overexpression vectors. (G) PCR detection of PKM2 mRNA level modulated by UCHL1 in different UBC cell line groups respectively transfected with UCHL1-specific shRNAs or overexpression vectors (n = 3).