Day 1: Transfection of Hek293 GFP LC3 cells

Hek293 GFP LC3 cells were reverse transfected with siRNA using Lipofectamine 2000 transfection reagent in 96 well plate format.

 

Day 3: Media change

Culture media was removed and fresh full media was added to alllow the cells recover from the transfection stress. Plates then incubate overnight at 37°C.

 

Day 4: EBSS induction and cell fixation

Plates were washed with PBS and then incubated EBSS+Leupeptin for 2 hours at 37C.

Cells were then fixed with 4% formaldehyde.

Day 5: Cell staining

Plates were stained with DAPI.

Images were collected on ArrayScan microscope: