Immunoblotting was performed as described previously [25 (link)]. Primary antibodies against AKT (Cell Signaling; 9272S, 1 : 1000, Frankfurt, Germany), phospho‐Akt S473 (Cell Signaling; 9271, 1 : 1000), PME‐1 (Santa Cruz Biotechnology; sc‐20086, 1 : 1000, Heidelberg, Germany), phospho‐PDHE1α S300 (Merck Millipore; ABS194, 1 : 1000, Darmstadt, Germany), cleaved PARP1 (Abcam; ab32064, 1 : 1000, Cambridge, UK), SV40 T Ag (Pab 108) (Santa Cruz Biotechnology; sc‐148, 1 : 1000), β‐actin (Sigma‐Aldrich; A1978 , 1 : 10 000), and GAPDH (HyTest; 5G4cc, 1 : 10 000, Turku, Finland). Secondary antibodies were purchased from LI‐COR Biotechnology, Bad Homburg, Germany or Dako (Agilent Technologies, Winooski, VT, USA. The histological methods were performed by the Histology core facility of the Institute of Biomedicine, University of Turku, Finland. Brain sections were stained with phospho‐ERK1/2 (Thr202/Tyr204) (Cell Signaling; #9101, 1 : 500) and Ki67 (Dako; M7240, 1 : 500). Slides were scanned using a Pannoramic 250 Flash scanner (3DHISTECH, Budapest, Hungary), and images were acquired using a slideviewer v2.6 (3DHISTECH).
Free full text: Click here
