2.5. LDH Release Assays

The Invitrogen^TM CyQUANT^TM LDH Cytotoxicity Assay Kit (Invitrogen, Waltham, MA, United States) was used to determine the LDH release of non-cancer cells treated with CT-p19LC, according to the manufacturer’s instructions. Briefly, 16HBE14o- and MCF10A cells were seeded at densities of 7.5 × 10⁴ and 4.5 × 10⁴ cells/well (three replicates), respectively, and left to adhere and grow overnight in the same conditions. The next day, the medium was collected and the cells were treated with the peptides (100 μM). After 48 h, the medium was collected and analyzed. Untreated cells were used as controls to compare the spontaneous LDH release and to normalize the data. Additional controls used were the maximum LDH activity release by lysing the cells with the lysis buffer provided in the kit, as well as the LDH positive control.