Figure 4.

Improved therapeutic correction by consensus AGT-RHEAM in a cell model of enzyme replacement (ERT). CHO-GO cells, which represent a model of PH1, were incubated with fusion proteins made up of a TAT sequence (that confers cell-penetrating ability) attached to the N-terminus of AGT or AGT-RHEAM (TAT-AGTs). Stably transfected CHO-GO cells (CHO-AGT) were used as positive controls, while untransfected cells (CHO-GO) and cells treated with AGT without the TAT peptide (AGT) were used as negative controls. Panels show kinetics of transduction for TAT-AGT-RHEAM (measured by western-blot), dose-response curves for TAT-AGT and TAT-AGT-RHEAM (measured by western-blot and % of specific activity with respect to CHO-AGT cells), metabolic rescue (measured by an indirect glycolate toxicity assay), and intracellular AGT activity expressed as % with respect to CHO-AGT cells. Experiments were performed essentially as described in [76]. Experimental details can be found in Appendix A.