title: Monitoring of autophagy is complicated--salinomycin as an example.
abstract: Monitoring of autophagy is challenging because of its multiple steps and lack of single befitting technique for a complete mechanistic understanding, which makes the task complicated. Here, we evaluate the functionality of autophagy triggered by salinomycin (anti-cancer stem cell agent) using flow cytometry and advanced microscopy. We show that salinomycin does induce functional autophagy at lower concentrations and such a dose is cell type-dependent. For example, PC3 cells show active autophagic flux at 10 μM concentration of salinomycin while murine embryonic fibroblasts already show an inhibition of flux at such doses. A higher concentration of salinomycin (i.e. 30 μM) inhibits autophagic flux in both cell types. The data confirms our previous findings that salinomycin is an inducer of autophagy, whereas autophagic flux inhibition is a secondary response.
authors.affiliation: Department of Clinical and Experimental Medicine (IKE), Division of Cell Biology and Integrative Regenerative Medicine Center (IGEN), Linköping University, Sweden.
authors.forename: Jaganmohan Reddy
authors.initials: JR
authors.lastname: Jangamreddy
authors.affiliation: Department of Medicine/Infectious Diseases, Case Western Reserve University, Cleveland, OH 44106, USA.
authors.forename: Soumya
authors.initials: S
authors.lastname: Panigrahi
authors.affiliation: Department of Clinical and Experimental Medicine (IKE), Division of Cell Biology and Integrative Regenerative Medicine Center (IGEN), Linköping University, Sweden; Department of Pathology, Pomeranian Medical University, Szczecin, Poland. Electronic address: marek.los@liu.se.
authors.forename: Marek J
authors.initials: MJ
authors.lastname: Łos
mesh_terms: D000818:Animals; D001343:Autophagy / Q000187:drug effects* / Q000502:physiology*; D058948:Cell Tracking / Q000379:methods; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D051379:Mice; D011714:Pyrans / Q000494:pharmacology*; D000095662:Polyether Polyketides
chemical_list: D011714:Pyrans; C010327:salinomycin; D000095662:Polyether Polyketides
keywords: Autophagic flux; GFP-LC3; Salinomycin; Vacuolization; mTandem GFP-RFP LC3; p62/SQSTRM1