Fig. 6

BTX-A promoted ubiquitin-degradation of SNAP23 by targeting NEDD4. a CHX was used to treat microglia following LPS and BTX-A. Expression of SNAP23 was measured by western blotting assay at 0, 2, 4, 6, 8 h later, respectively. The stabilization of SNAP23 was analyzed. b MG132 was used to inhibit ubiquitin-degradation of protein. Expression of SNAP23 was examined using western blotting assay. ^##P < 0.01 compared with Ctrl group, ^$$P < 0.01 contrasted with LPS group, and ^&&P < 0.01 compared with LPS + BTX-A group. c Western blotting assay was performed to ensure expression of SNAP23 in microglia. ^##P < 0.01 compared with Ctrl group, ^$$P < 0.01 contrasted with LPS group, and ^@@P < 0.01 compared with LPS + BTX-A group. d Co-IP assay was carried out to clarify the interaction between NEDD4 and SNAP23. IgG was served as control group. e HA-Ub and pcDNA-NEDD4 were co-transfected into HEK293T cells treated with or without MG132. Expression of SNAP23 and the level of SNAP23 ubiquitination were ensured using western blotting assay