Fig. 5.

Role of NEMO sumoylation in JCV infection and NEMO association with ATM. a Effect of JCV T-antigen on Sumoylation of NEMO. TC620 cells were transfected with expression plasmids for FLAG-tagged NEMO and/or T-Ag. Western blots were performed with antibody to T-Ag and anti-FLAG antibody to detect NEMO and sumoylated NEMO (arrowhead) as indicated. The quantified intensity of the sumoylated NEMO band is indicated in the histogram directly below the Western. The intensity of the sumoylated band as a percentage of total NEMO with and without T-Ag is presented in the lower right-hand part of the panel. b Physical association between ATM and NEMO. TC620 cells were transfected with expression plasmids for ATM and FLAG-NEMO. Whole cell extract (Input, lane 1) was immunoprecipitated with rabbit anti-ATM (lane 3). Nonimmune rabbit serum (NRS) was the negative control (lane 2). Western blot was with α-FLAG antibody. c In a reciprocal immunoprecipitation, whole cell extract from Panel b (Input, lane 1) was immunoprecipitated with mouse anti-FLAG (lane 3). Nonimmune mouse serum (NMS) was the negative control (lane 2). Western blot was with α-ATM antibody