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Angiosarcoma tissue sample immunostaining. Biopsie from patients suffering from angiosarcoma.

FoxO1 phosphorylation status correlates in angiosarcoma patient survival (Riddell et al 2018 Nat Commun). The survival time of the pSer218 FoxO1 negative group was significantly longer than that of the pSer218 FoxO1 positive groups.

Formaldehyde-fixed paraffin block patient tissue samples were cut into 5-7 μM sections then deparaffinized in xylene and rehydrated in a graded ethanol series. This was followed by antigen retrieval in sodium citrate buffer by boiling the slides for 15 min. The tissue was then blocked in 5% FCS + 0.1% Tween-20 for 30 min at room temperature followed by incubation with primary antibodies overnight at 4 degrees (anti-pSer218-FoxO 1:100; anti-VE-Cadherin 1:100; R&D Systems AF938) Tissue was then washed and secondary antibodies were applied.

Image provided courtesy of Dr. Masanori Nakayama Laboratory