Culturing Cell Lines and Viruses for Infection Studies

LLC-MK2 (ATCC, CCL-7) and A549 cells (ATCC, CCL-185) were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Corning) supplemented with 8% fetal calf serum (FCS; Life Technologies), 25mM HEPES (Gibco) and 50 μg/ml Gentamicin (Life Technologies). Primary normal human bronchial/tracheal epithelial (HPBE) cells (ATCC, PCS-300-010) were maintained in airway epithelial cell basal media (ATCC, PCS-300-030) supplemented with bronchial/tracheal epithelial cell growth kit components (ATCC, PCS- 300–040). A549 cells constitutively expressing Rab11-mRFP or Rab8-mRFP were established as described before for HeLa cells [13 (link)]. Briefly, A549 cells were transfected with the mRFP-Rab11a or mRFP-Rab8a cDNA in pCAGGS-HygR vector and hygromycin resistant cells were selected and grown in DMEM containing 8% FCS (DMEM-FCS). SeV (strain Enders), hPIV1 (strain C-35) and hPIV3 (strain C243, ATCC VR-93) were grown in LLC-MK2 cells in DMEM supplemented with 0.15% bovine serum albumin (DMEM-BSA) and acetylated trypsin at 2 μg/ml.