Facscan aria 3

Hoechst 33342 exclusion (side population) assays were carried out as previously described [30] to analyse cells overexpressing ABC transporters. Single‐cell suspension obtained from parental cell lines and melanospheres were stained with Hoechst 33342 (Sigma‐Aldrich) dye. As negative controls, verapamil (Sigma‐Aldrich) was used for maintaining the efflux channel closed, inhibiting the capacity to efflux Hoechst 33342 by cells. The bright fluorescent cells were measured by flow cytometry in Hoechst blue (440/40) and Hoechst red (695/40) of a FACScan Aria III (BD Biosciences) using FACSDIVA software from the CIC Scientific Instrumental Centre (University of Granada). Cells with the ability to efflux Hoechst 33342 were considered as the side population (SP).

Hoechst 33342 exclusion (Side Population) assays were carried out as previously described to analyze cells overexpressing ABC transporters. Melanospheres were stained with Hoechst 33342 (Sigma-Aldrich) dye. As negative controls, Verapamil (Sigma-Aldrich) was used for maintaining the efflux channel closed inhibiting the capacity to efflux Hoechst 33342 by cells [14 (link)]. The brightly fluorescent cells were measured by flow cytometry in Hoechst blue (440/40) and Hoechst red (695/40) of a FACScan Aria III (BD Biosciences) using FACS DIVA software from the CIC Scientific Instrumental Centre (University of Granada). Cells with the ability to efflux Hoechst 33342 were considered as the side population (SP) [14 (link)].