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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV100076	1/1	Mesocricetus auratus	NAY	(d)(U)C	Evdokimovskaya Y	2010	11%
Study summary Full title Secretion of the heat shock proteins HSP70 and HSC70 by baby hamster kidney (BHK-21) cells All authors Evdokimovskaya Y, Skarga Y, Vrublevskaya V, Morenkov O Journal Cell Biol Int Abstract The HSPs (heat-shock proteins) of the 70-kDa family, the constitutively expressed HSC70 (cognate 70- (show more...)The HSPs (heat-shock proteins) of the 70-kDa family, the constitutively expressed HSC70 (cognate 70-kDa heat-shock protein) and the stress-inducible HSP70 (stress-inducible 70-kDa heat-shock protein), have been reported to be actively secreted by various cell types. The mechanisms of the release of these HSPs are obscure, since they possess no consensus secretory signal sequence. We showed that baby hamster kidney (BHK-21) cells released HSP70 and HSC70 in a serum-free medium and that this process was the result of an active secretion of HSPs rather than the non-specific release of the proteins due to cell death. It was found that the secretion of HSP70 and HSC70 is independent of de novo protein synthesis. BFA (Brefeldin A) did not inhibit the basal secretion of HSPs, indicating that the secretion of HSP70 and HSC70 from cells occurs by a non-classical pathway. Exosomes did not contribute to the secretion of HSP70 and HSC70 by cells. MBC (methyl-beta-cyclodextrin), a substance that disrupts the lipid raft organization, considerably reduced the secretion of both HSPs, indicating that lipid rafts are involved in the secretion of HSP70 and HSC70 by BHK-21 cells. The results suggest that HSP70 and HSC70 are actively secreted by BHK-21 cells in a serum-free medium through a non-classical pathway in which lipid rafts play an important role. (hide) EV-METRIC 11% (30th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin NAY Focus vesicles Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (d)(U)C Protein markers EV: non-EV: HSP70/ HSC70 Proteomics no Show all info Study aim Other/Secretion of HSP70 and HSC70 Sample Species Mesocricetus auratus Sample Type Cell culture supernatant EV-harvesting Medium serum free Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 300 Characterization: Protein analysis Western Blot Antibody details provided? No Detected contaminants "HSP70/ HSC70"

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EV-TRACK ID	EV100076
species	Mesocricetus auratus
sample type	Cell culture
cell type	NAY
condition	NAY
separation protocol	(d)(U)C
Exp. nr.	1
EV-METRIC %	11