Fig. 1

Breast cancer-derived exosomes reshape metabolic characteristics in adipocytes and skeletal muscle cells. (a) Representative immunohistochemistry staining of CD36 and FATP1. The pictures also show positive staining for FATP1 and CD36 in breast cancer tissues located near stromal adipocytes. (b) Immunofluorescence staining for myosin heavy chain 1 (MYH1) after treatment 24 h. Scale bars represent 50 μm. The C2C12 in the CytoD group were treated with cytochalasin D (final concentration, 2 μg/ml) and 50 μg of exosomes purified from cancer-associated conditioned medium (CA-CM). (c) Mature adipocytes were stained with red oil after treated by CytoD and/or exosomes. The adipocytes in the CytoD group were treated with cytochalasin D (final concentration, 2 μg/ml) and 50 μg of exosomes purified from cancer-associated conditioned medium (CA-CM). (d) The levels of secreted metabolites (pyruvate, lactate, glycerol and free fatty acids) enriched in media were determined by colorimetric assay. AD: adipocytes. (e) Raw data for the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) as determined by the Seahorse XF24 analyser. The ECAR was evaluated after the addition of 10 mM glucose to adipocytes or C2C12 in the presence or absence of exosomes. The OCR was measured in the presence of palmitate as described in the Methods. (f) Adipocytes or C2C12 were cocultivated in the presence or absence of exosomes. The proteins were extracted for western blot analysis of the expression of the indicated proteins. Data are presented as the mean ± S.D. of at least three independent experiments. * P < 0.05 versus control values, # P < 0.05 versus control values as positive group