Figure 1
From: A new synthetic protein, TAT-RH, inhibits tumor growth through the regulation of NFκB activity
Effects of GRK5-RH in vitro in KAT-4 cells overexpressed by adenovirus mediated gene transfer. A) IκBα and GRK5-RH levels were analyzed in whole extracts by western blot, in KAT-4 overexpressing GRK5-RH by means of adenovirus mediated gene transfer. GRK5-RH increases IκBα levels. GRK5-RH expression was visualized by WB using anti-GFP antibody. B) To evaluate GRK5-RH effect on apoptosis, we analyzed the cleavage of caspase 3 by WB. The overexpression of GRK5-RH increases cleaved caspase 3 levels, suggesting that GRK5-RH causes an increase of apoptotic responses. C) This result was confirmed by Annexin V staining in fluorescence. GRK5-RH overexpression causes apoptosis as shown by Annexin-V staining compared to live cells (green = Annexin V; blue = nuclei) D) We evaluated GRK5 effects on NFκB activity by luciferase assay, in KAT-4 overexpressing GRK5-RH. Cells were transfected with plasmids coding for a κB-luciferase reporter (κB-Luc) and β-galactosidase (β-Gal) and lysates were analyzed by luciferase assay system (Promega). GRK5-RH inhibits NFκB transcriptional activity (*p < 0.05 vs control).