Fig. 4

USP22 silencing antagonized the effect of KLF3-AS1 on Sirt1 deubiquitination. A, B BV-2 cells were transfected with KLF3-AS1 overexpression plasmid or vector, Sirt1 ubiquitination was evaluated by Co-immunoprecipitation (A), then protein expressions of USP22, MDM2 and P-JNK were tested (B). Cells were transfected with pcDNA-KLF3-AS1 or sh-USP22, then Sirt1 and USP22 proteins were detected by western bolt (C), and Sirt1 deubiquitination was assessed by Co-immunoprecipitation (D). E Cells were transfected with KLF3-AS1 overexpression plasmid or vector, the interaction between USP22 and Sirt1 was then validated by Co-immunoprecipitation. Values were expressed as mean ± SD of three separate determinations in the in vitro assays. **P < 0.01, ***P < 0.001