HLMs were cultured in 24-well plates in RPMI 1640 medium supplemented with 5% FCS, 2 mM l-glutamine, and 1% antibiotic–antimycotic solution, as previously described [17 (link)]. HLMs were treated with TSLP and its proteolytic fragments for 16 h at 37 °C. At the end of incubations, the supernatants were collected and stored at −80 °C for subsequent ELISA quantification of VEGF-A. Cell lysis in the plates was carried out using 0.1% Triton X-100 for total protein quantification by a Bradford-based assay (Bio-Rad, Segrate, Milan, Italy).
Canè L., Poto R., Palestra F., Iacobucci I., Pirozzi M., Parashuraman S., Ferrara A.L., Illiano A., La Rocca A., Mercadante E., Pucci P., Marone G., Spadaro G., Loffredo S., Monti M, & Varricchi G. (2024). Thymic Stromal Lymphopoietin (TSLP) Is Cleaved by Human Mast Cell Tryptase and Chymase. International Journal of Molecular Sciences, 25(7), 4049.
Other organizations :
University of Naples Federico II, World Health Organization - Italy, Institute for Experimental Endocrinology and Oncology, National Research Council, Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale"
HLMs were cultured in 24-well plates in RPMI 1640 medium supplemented with 5% FCS, 2 mM l-glutamine, and 1% antibiotic–antimycotic solution
Cell lysis in the plates was carried out using 0.1% Triton X-100 for total protein quantification by a Bradford-based assay
controls
No positive or negative controls were explicitly mentioned.
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