<p>(A) BJAB and HEK-293 cells were co-transfected with constructs pCDNA3.1-HAChk2 and pA3M-LANA. Co-immunoprecipitation from the cell lysate was performed by using anti-Myc antibodies. The co-immunoprecipitates were separated by electrophoresis, transferred to a nitrocellulose membrane, and then probed with HA antibodies for Chk2. Chk2 immunoprecipitated with LANA in both cell types. (B) BJAB cells were co-transfected with the expression constructs pCDNA3.1-HAChk2 and pA3M-LANA. Following transfection, the cells were grown overnight and fixed. LANA and Chk2 were detected by using mouse monoclonal antibody against Myc-LANA and rabbit polyclonal antibody against HA- Chk2, followed by appropriate secondary antibodies conjugated to Alexa Fluor 488 (green) and Alexa Fluor 594 (red), respectively. The merged panel shows that Chk2 and LANA co-localize in the nucleus. The DAPI panel shows that both proteins are nuclear. (C) Schematic representation of full-length domains along with the different truncation constructs of Chk2. FHA: fork head association domain. (D, E) <i>In vitro</i> translated LANA or KSHV-positive BC3 cells nuclear extract were incubated with the various GST-Chk2 truncated constructs as shown in figure. The pull-down assay showed a preferential binding for the region located between amino acids 63 and 107, which includes the serine rich domain. NE, nuclear extract.</p
