Fig. 7
A20 upregulated STC1 by inhibiting STC1 degradation. a, b Degradation of STC1 protein over time in cells with or without A20 overexpression (OE). Cycloheximide was used at 20 μM. c The interaction between A20 and STC1 detected by Co-IP. d The co-localization of STC1 (green) and A20 (red) (×1000) detected by immunofluorescence assay. Blue dye, DAPI indicates the nucleus, MitoTracker (white). e Schematic diagram of A20 domains in the various constructs. f, g The interaction between STC1 and A20 with different ___domain-deletions. h The effect of A20 with different ___domain-deletions on STC1 expression. i Schematic diagram showing 4 STC1 expression vectors with different C-terminal deletions. j The interaction between A20 and STC1 (having different C-terminal deletions). k The amino acid sequence of STC1 from 60 to 88aa among different species. l Degradation profile of wild-type STC1 protein and STC1 with T86A mutant (0, 0.5, 1, and 2 h). m The effect of STC1 T86A or T86D mutant on its binding to A20. n The interaction between STC1 and GSK3β. o The co-localization of GSK3β (green) and STC1 (red) (×630) detected by immunofluorescence assay. Blue dye, DAPI indicates the nucleus. p–r The effect of GSK3β on STC1 protein degradation and expression. s The effect of STC1 T86A or T86D mutant on the binding to GSK3β. t, u The effect of A20 on the interaction between STC1 and GSK3β. v The effect of MG132 (20 μM) and chloroquine (40 μM) on STC1 expression. w The effect of A20 on ubquitination of STC1 protein in 293T cells. Whole cell lysates were harvested after incubation with MG132 (30 μM for 12 h)
