Journal: PLoS ONE
Article Title: Proteasome Inhibition Contributed to the Cytotoxicity of Arenobufagin after Its Binding with Na, K-ATPase in Human Cervical Carcinoma HeLa Cells
Figure Lengend Snippet: Cellular proteasomal activity was inhibited by arenobufagin, which might be related to the binding of Na, K-ATPase. (A ) Inhibition of arenobufagin (Arg) at different doses on cytosolic proteasomal activity for 24h. ( B ) Three types of cellular proteasome enzyme activities (C-L, T-L and CT-L) in HeLa cells treated with 0.1% DMSO or arenobufagin(Arg) at different concentrations for 24h. ( C ) Western blotting results of WEE1 and actin in control HeLa cells and cells treated with 10 nM arenobufagin for 24 h and 48 h. Each blot was the representative result of three independent experiments. ( D ) Inhibition of antibodies against α1 or α3 subunits of Na,K-ATPase or combination of antibodies against α1 or α3 subunits of Na,K-ATPase and 10 nM arenobufagin on cytosolic proteasomal activity for 24h,48h and 72h, respectively. Data were statistical results of three independent experiments. Data were statistical results of three independent experiments. Data were expressed as mean ± SD. *Significant difference from the control group at P
Article Snippet: The primary antibodies used in the present study were mouse anti-Na, K-ATPase α1 monoclonal antibody (1:200, Santa Cruz Biotechnology), mouse anti-Na, K-ATPase α3 monoclonal antibody (1:200, Santa Cruz Biotechnology), mouse anti-WEE1 monoclonal antibody (1:500, Abnova, Taipei City, Taiwan), mouse anti-GAPDH monoclonal antibody(1:2000, Sigma) and mouse anti-actin monoclonal antibody (1:2000, Sigma).
Techniques: Activity Assay, Binding Assay, Inhibition, Western Blot