Figure 2.
Expression of full-length ATM cDNA in A-T cells. EBV-transformed lymphoblastoid cells were transfected with pMAT1 by using the Lipofectin reagent (Life Technologies) according to the manufacturer’s instructions. Stably transfected A-T cells were induced with 5 μM CdCl2 (+) for 16 h or were uninduced (−). Total cell extract from pMAT1 transfected AT1ABR and AT3ABR were immunoblotted with different ATM antibodies. A control cell line, C3ABR, and an A-T cell line, L3 (homozygous mutations at nucleotide 120 predicted to give rise to truncated protein 35 amino acids long), are also included. (a) ATM-1.8 antibodies. (b) ATM4BA antibodies. (c) Analysis of samples purified from cell extracts by using a nickle chelate resin to isolate His-tagged protein. Detection was carried out with ATM4BA antibodies. (d) Analysis of samples immunoprecipitated with anti-hexahistidine antibody (CLONTECH) followed by immunoblotting with ATM4BA antibody.