FIG 2.

JEV infects and replicates in porcine NEC. In panel A, porcine NEC cultures were infected with JEV strains at MOI of 0.1 TCID₅₀/cell, and after 24, 48, and 72 hpi, inserts were collected and viral RNA was quantified by real-time RT-PCR. In panels B and C, viral titers from the apical and basolateral compartments of the same cultures as in panel A are shown. In panels D and E, JEV-infected porcine NEC were analyzed by multicolor immunostaining for nuclei (DAPI, dark blue), cilia (β-tubulin, red), tight junctions (ZO-1; light blue), and JEV E protein. 3D scans were acquired using confocal microscopy; scale bars represent 20 and 8 µm for panels D and E, respectively. The experiment was repeated three times in duplicate. In panels A to C, results are represented as 25 to 75% interquartile boxes showing the means and 95% confidence intervals. Different superscript letters indicate significant differences (P < 0.05) between samples challenged with the same JEV strain (letters without and with apostrophe for the Laos and Nakayama strains, respectively). Significant differences between distinct JEV strains at the same time postinfection are shown by an asterisk for virus titers (A) and for RNA levels (B and C).