Figure 2.

Generation of an inducible AGAL and mutant rescue in Fabry podocytes. (A) Schematic overview of the doxycycline inducible pINDUCER construct (tet on—tet off) driving wild-type AGAL expression after stable lentiviral-mediated genomic insertion. (B) AGAL activities in wild-type, AGAL-deficient and lentiviral-mediated transfected podocytes with and without doxycycline induction. (C) Western blot analysis of AGAL expression in lentiviral-mediated rescued wild-type doxycycline-induced podocytes. AGAL: α-galactosidase A, Dox (-): without doxycycline, Dox (+): with doxycycline, Ef1a: elongation factor 1 alpha, IRES: internal ribosomal entry site, LTR: long terminal repeat, Puro: puromycin resistance, rtTA: reverse tetracycline-controlled transactivator, TRE: tetracycline response element, WT: wild-type.