Figure 3.

ANP32B contributes to influenza A virus replication in human cells. (A,B) Viral titers in H3N2 or H5N1 infected human HeLa cells deficient for ANP32B were determined by plaque test at the indicated time points. Two different knockout cell lines were used to exclude clonal artifacts (ANP32B KO #1 and ANP32B KO #2, respectively). (C) Knockout of ANP32B in two clonal HeLa cell lines was verified by Western blotting. (D,E) Expression of viral nucleoprotein (NP) in H3N2 infected HeLa cell lines deficient for ANP32B was analyzed by Western blotting at 24 h p.i. Quantification of viral NP expression from two independent experiments was performed using ImageJ software (E). (A–E) Statistical significance was calculated using the two-tailed Student's t-test (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001; ns, not significant).