Inlab nano

Colorectal cancer cells HCT116, HT29, RKO were routinely cultured in DMEM culture medium supplemented with 10% FCS (HyClone Laboratories) and gentamicin (Sandoz). Experiments in hypoxia (1% O₂) were done in a humidified anaerobic workstation (Ruskinn Technologies) with 5% CO₂, 10% H₂ and 84% N₂ at 37 °C. All cell lines were purchased from ATCC and regularly tested for Mycoplasma contamination. Cells were treated with beta-blocker propranolol dissolved in DMSO in the final concentration 50 µM. Control cells were cultivated in the presence of an appropriate volume of DMSO. At the end of each experiment, pH of the culture medium was immediately measured by pH meter Seven2Go, S2, Mettler Toledo and microelectrode InLab Nano (Switzerland). The pH values were subsequently normalized to total protein concentration of each sample.

The concentration of lactate was determined in the whole body of E. verrucosus using the express kit “Lactate-vital” (Vital-Diagnostics, St. Petersburg, Russia) according to Bergmeyer47 . Measurements were performed with the “Cary 50” spectrophotometer (Varian, USA) at λ = 505 nm.
Haemolymph pH of E. verrucosus at acclimation conditions was measured by a microelectrode (InLab Nano, Mettler Toledo) at approximately 6 °C right after haemolymph extraction.