α pchk2

The cells were homogenized in lysis buffer (50 mM/L HEPES pH 7.5, 150 mM/L NaCl, 10% glycerol, 1% Triton X-100, 1 mM/L EGTA, 1.5 mM/L MgCl₂, 10 mM/L NaF, 10 mM/L sodium pyrophosphate, 1mM/L Na₃VO₄, 10 μg approtinin/ml, 10 μg leupeptin/ml) (Sigma). The following antibodies were used: α-RAS, α-RAC, α-CDC42, α-RHO, α-pEGFR, α-EGFR, α-pFAK, α-FAK, α-pSCR, α-SCR, α-pMEK, α-MEK, α-pERK1/2, α-ERK1/2, α-pAKT, α-AKT, α-pGSK3 α /β, α-GSK3 α /β, α-β catenin, α-pCHK2, α-CHK2, α-tubulin, α-SP1, α-pRET, α-RET (Cell Signaling, Danvers, MA, USA), and α-SOD3 (Santa Cruz, Santa Cruz, CA, USA). Signal density analysis was performed using ImageJ software GEL blot software.

Eyes and teratomas were removed immediately after euthanasia and fixed in 4% formaldehyde for at least 24 hr. For histological analysis, formaldehyde fixed tissues were embedded in paraffin, cut into 5 μm sections and stained with Hematoxilin and Eosin. The antibodies used were α-p53 (VectorLabs), α-p-ATM (Cell signaling), α−γ−H2AX (Cell signaling), α-p-CHK2 (Cell signaling) and α-p-ATM (Genetex).