Figure 1. Intracytoplasmic soluble expression of intrabody-PEST and intrabody-SCRPEST constructs is increased significantly with respect to intrabody alone. (A) D5E scFv, designated E; (B) 10H scFv, designated H; (C) D10 scFv, designated D; (D) VH14, designated V. ST14A cells were transiently transfected with Intrabody-hemagluttinin (HA), Intrabody-HA-PEST or Intrabody-HA-SCRPEST constructs. 48 h post-transfection, cells were harvested, cell lysates prepared, soluble protein separated and transferred on western blots. Proteins were identified using anti-HA monoclonal antibodies, with actin as a loading control. Proteins were quantified densitometrically and normalized to actin. Negative control lanes, empty vector only (pcDNA3.1-), were always blank. At least 3 independent experiments were performed and representative gels are illustrated. One-way ANOVA with Minitab statistical software was used to perform statistical significance. (*p < 0.05, **p < 0.01, compared with intrabody-HA)