Skip to main content
Fig. 4 | Molecular Medicine

Fig. 4

From: Heat-shock protein 90α is involved in maintaining the stability of VP16 and VP16-mediated transactivation of α genes from herpes simplex virus-1

Fig. 4

VP16 interacts with Hsp90α via its conserved core domain. a, b Hsp90 inhibition led to VP16 degradation in FLAG-VP16 plasmid-transfected cells. SH-SY5Y cells (a) and Vero cells (b) were transfected with FLAG-VP16 plasmid (3 μg) for 48 h and then treated with AT533 (2 μM) for 2 h. Proteins were extracted and subjected to western blot analysis; P3*FLAG-CMV was included as an empty vector control. c Hsp90α knockdown contributed to VP16 degradation in VP16-expressing cells. SH-SY5Y cells were cotransfected with siHsp90α-2 (100 nM) and FLAG-VP16 plasmid (3 μg) for 48 h. The cell lysates were then analyzed by western blotting. d SH-SY5Y cells were infected with HSV-1 (MOI 50) for 2 h in the presence of AT533 (2 μM), immunoprecipitated with anti-VP16 antibodies, and subjected to western blot analysis using anti-Hsp90 antibodies. Oct-1 was used as a positive indicator that interacts with VP16. e VP16 interacted with Hsp90α in 293 T cells. 293 T cells were cotransfected with FLAG-VP16 (5 μg) and HA-Hsp90α (5 μg) plasmids for 48 h. The cells were then lysed, subjected to immunoprecipitation using anti-FLAG antibodies, and then analyzed by western blotting. f Diagrammatic sketch of VP16 truncated mutant construction. g VP16 interacted with Hsp90α via its conserved core domain. 293 T cells were cotransfected with different truncated mutant plasmids of VP16 (5 μg), including WT, V1, V2, and V3, together with HA-Hsp90α (5 μg) plasmid for 48 h. The cells were lysed and subjected to immunoprecipitation using anti-FLAG antibodies and then analyzed by western blotting

Back to article page