Intracellular HRV RNA load in CF HBE cells after a short-term inoculation with HRV16.

<p>Primary CF HBE cells were pre-incubated for 18 hours with experimental culture medium (BEGM without hydrocorticortisone or antibiotics) and subsequently inoculated for 2 hours with HRV16 (HRV) at an MOI of 0.1 or 1.0. Then, virus-containing cell supernatants were removed and cells were rinsed five times with experimental culture medium to deplete extracellular virus. Subsequently, cells presented in left panels were fixed with paraformaldehyde and processed as described in the Materials and Methods to detect the HRV16 positive strand RNA by <i>in situ</i> hybridization. Parallel cells (right panels) were incubated for 2 hours post-inoculation (p.i.) and processed similarly to above cells. Green indicates expression of <i>ACTB</i> (β-actin gene; used to visualize cell cytoplasm), red dots and white arrows indicate the HRV16 positive strand RNA, and blue is DAPI (nuclear counterstain). The red and blue signals have been slightly overexposed to better visualize HRV16 RNA.