Open Access Highly Accessed Research article

Hypoxic enhancement of exosome release by breast cancer cells

Hamish W King12, Michael Z Michael2 and Jonathan M Gleadle1*

Author Affiliations

1 Renal Department, Flinders Medical Centre, Flinders University School of Medicine, Bedford Park, South Australia, 5042, Australia

2 Department of Gastroenterology and Hepatology, Flinders Medical Centre, Flinders University School of Medicine, Bedford Park, South Australia, 5042, Australia

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BMC Cancer 2012, 12:421  doi:10.1186/1471-2407-12-421

Published: 24 September 2012

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Additional File 1 :

Impact of hypoxia on cell growth and viability. (A, B) MCF7, SKBR3 and MDA-MB 231 breast cancer cells were cultured for 48 hours under normoxia or 1% O2. Cell counts were performed for each well after hypoxic exposure (A) and cell viability was determined by Trypan blue exclusion (B) (n=4; ± SEM). (C, D) MCF7, SKBR3 and MDA-MB 231 breast cancer cells were cultured for 24 hours under normoxia or 0.1% O2 and cell counts (C) and viability (D) data were obtained as described above (n=4; ± SEM). ** corresponds with P value < 0.01. (PDF 13 kb).

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Additional File 2 :

-isolated exosomes. Representative video of MCF7 exosomes isolated by ExoquickTM precipitation as visualised by Nanosight LM10 microscope using a 405 nm laser. (MPG 2468 kb).

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Additional File 3 :

Hypoxic enhancement of exosome release as detected by CD63 immunoblot. (A) CD63 and CD9 immunoblot of SKBR3 ExoquickTM precipitants from a 48 hour culture under normoxia or 1% O2, including band intensity quantitation. (B) CD63 immunoblot of SKBR3 ExoquickTM precipitants from a 24 hour culture under normoxia or 0.1% O2, including band intensity quantitation. All CD63 immunoblots were performed under non-reducing conditions as described previously [16]. (PDF 29 kb).

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