Fig. 4

USF1 transcriptionally activated USP14 that caused NLRC5 de-ubiquitination. (A) Prediction of USF1 binding motif in the promoter of USP14 using the JASPAR database. (B) Relative luciferase activity in HUVECs transfected with luciferase plasmid constructs containing the USP14 promoter (wt and mut) in combination with USF1 overexpression plasmid. (C) ChIP assay confirmed the binding of USF1 to the USP14 promoter. (D) HUVECs were transfected with sh-USF1 or oe-USF1. USF1 and USP14 levels were measured by RT-qPCR. (E) Ubibrowser2.0 database predicted the potential de-ubiquitination targets of USP14. (F) Co-IP assay analysis of NLRC5 ubiquitination. (G) USP14 and NLRC5 levels were determined by RT-qPCR and Western blotting. n = 3. *p < 0.05, **p < 0.01, ***p < 0.001