HBEC30-KT cells [34] were plated in 96 well plates and incubated overnight. Cells were infected with WSN virus at an MOI 5. Whole cell lysates were collected at the indicated time point and separated by 12% SDS-PAGE gel and transferred to a nitrocellulose membrane. Cultures for immunofluorescence were fixed with 4% formaldehyde at indicated time point. Viral protein was detected in both cases with antibodies for pan influenza A (1:200), M2 (1:500) or NP (1:500) proteins{a-tubulin (cellsignalling rabbit mAb, cat#2125S), anti-NS1 [35] , anti-NP (Abcam, cat# ab20343)} followed by detection with either HRP conjugated secondary or staining with Alexa 498 (1:5,000) or Alexa 594 (1:5,000) conjugated secondary antibodies(from Invitrogen). Wells were imaged with a 20x lens on a BD Pathway 855 microscope. Imaged cells were segmented using Hoescht staining and distance from nucleus, aIVA fluorescence intensity was measured, with Attovision software.