Figure 2
Phage EK99P-1 restored the decreased expression of tight junction proteins by ETEC in IPEC-J2 infected with ETEC K99. A monolayer of confluent IPEC-J2 was treated with ETEC K99 (2 × 106 cfu/mL) and phage EK99P-1 (2 × 105 pfu/mL) for 3 h. (A) Whole-cell lysates were analyzed for the expression of ZO-1, occludin, claudin-3, and β-actin using anti-claudin-3, -occludin, -ZO-1 and -β-actin antibodies by western blot assay (n = 3). Full-length blots/gels are presented in Supplementary Fig. 4. (B) ZO-1 expression in IPEC-J2 was visualized using confocal microscopy after staining with anti-ZO-1 antibody conjugated with Alexa Fluor 488-FITC (green) and nuclei with DAPI (blue) (n = 3). Scale bar = 30 μm.
