Modfit software v 5

The analysis of the cell cycle in HRECs and iCEC2 cells was assessed as described before^{38 (link)}. Briefly, cells were grown in 6 well plates (2 × 10⁶) and at 70% confluency, the cells were treated with PR5-LL-CM01 (0.1 µM, 1 µM and 10 µM) or vehicle DMSO (1% final concentration to the cells) for 24 h. The cells were washed with PBS twice and fixed in 70% ethanol. Prior to analysis, the cells were washed with PBS for two times, the cell pellets resuspended in propidium iodide (PI) staining solution (20 µg/ml) prepared in 1× PBS containing 100 µg/ml of RNAse A and 0.1% Triton X-100 and incubated at room temperature for 30 min. The cells were then analyzed on an Attune NxT flow cytometer (Thermo Fisher Scientific, Waltham, MA, USA). The single cell population was analyzed by area histograms and cell cycle profiles were created using Modfit software (v. 5.0, Verity Software House, Topsham, ME, USA). Pulse shape analysis was performed to eliminate any debris, doublets and aggregates from the whole cell population analyzed.