Transfection by electroporation of GFP-LRRK2 and Immunofluorescent imaging of MEFs VPS35 (D620N) mutants stably expressing LysoTag

Transfection of foreign DNA and Immunofluorescent (IF) microscopy are powerful tools used in cellular and molecular biology to monitor subcellular localisation of proteins. Mouse embryonic fibroblasts (MEFs) are primary cells notorious for their low transfection efficiency by mostly available chemical methods. This efficiency become even lower if the aim is to express a large sized protein, such as LRRK2, a 250kD protein. Here, we described a method where we used electroporation to transiently transfect GFP-tagged LRRK2 into MEFs. We also used IF microscopy to visualise the subcellular localisation of the transiently expressed GFP-LRRK2. Furthermore, we investigated the colocalization of GFP-LRRK2 with a lysosomal localised TMEM192-3xHA.