4.6. Cell Proliferation Assay

The proliferative status of fibroblasts was analyzed using the Click-iT EdU Alexa Fluor 488 Flow Cytometry Assay Kit (Invitrogen) according to the manufacturer’s instructions. Briefly, cultured fibroblasts were treated with or without IL-1β (10 ng/mL) for 24 h before the addition of the nucleoside analog EdU (5-etinil-2´-deoxiuridin), which is incorporated into the DNA only by cells under replication. Proliferative cells were evaluated using a FACSCanto II Cytometer (BS Biosciences) and BD FACSDiva software v.6.0 software (San Jose, CA, USA). Results are shown as % Click-iT^® positive cells (proliferating cells) with respect to total cells.