Standard diluent

Tumor cells were grown in transwell with adipocytes in the presence or absence of 10 μM Atglistatin. After 48 hours, adipocytes were collected as previously indicated and re-suspended in the Standard Diluent (provided in the Triglyceride assay kit; Cayman Chemical). Samples were sonicated and centrifuged. The supernatant was then used for the assay. All steps were performed according to manufacturer's protocol. Experiments were done in triplicate and reported as mean ± SD.

Frozen liver was minced into pieces and homogenized in standard diluent (Cayman Chemical, Ann Arbour, Michigan, USA) with 20 µg/mL Leupeptin (Serva Electrophoresis GmbH, Heidelberg, Germany) via a rod-homogenizer for 10–15 s. Homogenates were centrifuged at 4 °C for 10 min at 10,000× g. Supernatant was aliquoted for triglyceride and protein quantification. Colorimetric detection and quantitation of total protein was performed using the Pierce™ BCA Protein Assay Kit (Thermo Fisher Scientific Waltham, Waltham, MA, USA). Triglyceride in supernatant was detected via the Cobas^® 8000 modular analyzer (HITACHI/Roche, Basel, Schweiz).