For preparation of AnExILs with b-galactosidase expressed inside liposomes (further referred to as AnExIL-IN), 75 ll of IVTT mixture and pIVEX2.2EM-LacZ with a final concentration of 20 nM, was used to rehydrate a batch of 6 lM of PEG-lipid cakes in order to form liposomes encapsulating IVTT mix and pDNA. The liposomes were incubated on ice for 10 min to complete the rehydration process. To inactivate protein expression outside liposomes, RNase with a final concentration of 10 lg/ml, was added to the liposomal suspension. Samples were incubated at 30°C for 3 h to allow protein synthesis to complete.