2.4 | In vitro cell membrane injury assay
We performed an in vitro cell membrane injury repair assay following the
protocols described previously.26, 29 HNECs were
treated with TGF-β1/TGF-β1 inhibitor or transfected with Ad-MG53 for 72
h, and then suspended in PBS. Next, 100 μL of the cell suspension (5 ×
104 cells/well) was added to a 96-well plate along
with acid-washed glass microbeads. The plate was shaken at 200 rpm for 5
min to induce cell membrane injury, followed by centrifugation at 3000
rpm for 10 min, and 80 μL of the supernatant was collected. Lactate
dehydrogenase (LDH) activity in the supernatant was determined using the
LDH Cytotoxicity Detection Kit (MeiKe, MK0354A, China).