Live Cell Imaging of p62 using LentiBrite™ Fluorescent Biosensors

Human mesenchymal stem cells (MSCs), were plated in a chamber slide and transduced with LentiBrite™ RFP-p62 lentiviral particles at an MOI of 40 for 24 hours. After media replacement and 24 hours further incubation, cells were incubated for 4 hours in EBSS containing a lysosome inhibitor, to induce autophagy and inhibit lysosomal degradation of autophagosomes. The RFP-p62 displays a diffuse cytosolic distribution with scattered small puncta in fed cells, and an entirely punctate distribution in starved cells. LentiBrite™ Lentiviral Biosensors are a new suite of pre-packaged lentiviral particles encoding important and foundational proteins of autophagy, apoptosis, and cell structure for visualization under different cell/disease states in live cell and in vitro analysis.

Materials