Detection of the RV RNA-Calreticulin Complex in Infected
Vero 76 Cells. Uninfected, RV-infected, and RV-infected
cells transiently expressing calreticulin fused to a FLAG
peptide (IBI/Eastman Chemical) (C.D.A., unpublished data)
were exposed to UV irradiation on ice-water for 30 min prior
to lysis. Cytolysates were prepared and subjected to immunoprecipitation
using anti-calreticulin and anti-FLAG antibodies
as described (7). In a control experiment, the uninfected
cell lysate was UV-crosslinked with a synthetic RNA
corresponding to nt 3220-3381 of RV RNA (16). The immunocomplexes
were eluted in 100 /A of sterile water at 100°C
and treated with proteinase K (100 ng/,ul) in the presence of