flow cytometry assays can be employed. However, the presence of cytosolic LC3-I (non-lipidated form) can cause persistent background signal and can make quantitative analysis difficult via flow cytometry.
In this chapter, we describe the utility of this probe for measuring autophagic activity in cultured cells by flow cytometry and for analyzing autophagic activity in zebrafish tissues by confocal microscopy.doi:10.1007/978-1-0716-1589-8_4Tomoya Eguchi...
flow cytometry demonstrated theexpression rates of RFP and GFP reached to 100%. The number of autophagic puncta significantly increased after starvationtreatment. Conclusion The RAW264.7 cell strain with stable expression of RFP-GFP-LC3 has been successfully constructed,which provides a reliable cellular...