Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining of HPAECs was performed using a TUNEL kit (Cat# C10617, Thermofisher) as per manufacturer instructions. Five randomly selected fields were photographed, and the number of apoptotic nuclei as well as the total number of nuclei was counted per HPF. The apoptotic index was obtained by means of the formula: number of apoptotic cells per field / total number of cells per field as previously described31 (link).
Batlahally S., Franklin A., Damianos A., Huang J., Chen P., Sharma M., Duara J., Keerthy D., Zambrano R., Shehadeh L.A., Martinez E.C., DeFreitas M.J., Kulandavelu S., Abitbol C.L., Freundlich M., Kanashiro-Takeuchi R.M., Schmidt A., Benny M., Wu S., Mestan K.K, & Young K.C. (2020). Soluble Klotho, a biomarker and therapeutic strategy to reduce bronchopulmonary dysplasia and pulmonary hypertension in preterm infants. Scientific Reports, 10, 12368.
Apoptotic index - calculated as the number of apoptotic cells per field divided by the total number of cells per field
control variables
Five randomly selected fields were photographed
controls
No positive or negative controls were explicitly mentioned
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