Edoardo Bellini

Abstract Why some tumors respond to immunotherapy (“hot” tumors) while others remain resistant (“cold” tumors) is a central challenge in oncology. Elevated RAB5A-dependent endocytosis drives tissue fluidization during the transition to invasive breast carcinoma, but its immunological consequences are unclear. Here we show that RAB5A-driven fluidization induces a mechano-metabolic stress response that disrupts the AMPK–AKAP1–DRP1 mitochondrial fission pathway, causing mitochondrial elongation.

Quantitative RT-PCR Analysis Quantitative RT-PCR analysis was performed as previously described.[3] Total RNA was extracted using the RNeasy Mini kit (Qiagen) and quantified by NanoDrop to assess both concentration and quality of the samples. 1 microgram of RNA was retrotranscribed using “High-Capacity cDNA Reverse Transcription Kit – Thermofisher.” RT minus was made to check the absence of genomic DNA.

Abstract Untargeted metabolomics by mass spectrometry technologies generates huge numbers of metabolite signals, requiring computational analyses for post-acquisition processing and databases for metabolite identification. Web-based data processing solutions frequently include only a part of the entire workflow thus requiring the use of different platforms.