Next, we tested the cellular localization of truncated NSs other than NSD249-265 by Western blot (Fig. 4) . We did not include NSD249-265 for the experiment as no antibodies were available to detect this NSs in Western blot. 293 cells were mock-infected or infected with MP-12 or NSs truncation mutants at an moi of 3. Cells were collected at 16 hpi, and nuclear and cellular fractions were analyzed for the presence of NSs proteins. MP-12 NSs were accumulated both at cytoplasm and nucleus, while N proteins were exclusively localized at cytoplasm, which is consistent with previous study [65] . Abundant accumulation of NSs in nucleus was only observed in cells infected with NSsD6-30, NSsD 31-55 and NSsD56-80, while other mutants, NSsD 81-105, NSsD106-130, NSsD131-155, NSsD156-180, NSsD181-205 and NSsD206-230, NSsD231-248 poorly accumulated NSs in nucleus.