Nicholas J. Rossiter

ArticleVolume 38, Issue 8p1599-1617.e8Open access Affiliations & Notes 1University of Michigan, Department of Molecular & Integrative Physiology, Ann Arbor, MI, United States of America 2University of Michigan, Department of Biological Chemistry, Ann Arbor, MI, United States of America 3University of Michigan, Program in Cellular and Molecular Biology, Ann Arbor, MI, United States of America 4Medical Scientist Training Program, University of Michigan, Ann Arbor, MI, 48109, United States of...

Results To visualize ERMCSs in living cells, we generated eleven isogenic cell lines expressing a doxycycline-inducible split-GFP-based ERMCS reporter (SPLICS) together with a mitochondrial matrix red fluorescent protein reporter (mitoTagRFP)14,15,16,17,18 (Figure 1A). Selected clones exhibited inducible and reversible GFP fluorescence with stable RFP expression. Reporter induction did not affect cell growth, oxygen consumption, or extracellular acidification (Figures S1A–S1C).

Abstract Pancreatic ductal adenocarcinoma (PDAC) subsists in a nutrient-deregulated microenvironment, making it particularly susceptible to treatments that interfere with cancer metabolism1,2. For example, PDAC uses, and is dependent on, high levels of autophagy and other lysosomal processes3,4,5. Although targeting these pathways has shown potential in preclinical studies, progress has been hampered by the difficulty in identifying and characterizing favourable targets for drug development6.