FIG. 5.
Comparison of parental Rov9 cells and the most SSBP/1-permissive first- and second-generation subclones. (a) Dose-response graph showing the average number of infected cells (spots) detected by the SSCA when cells were challenged with increasingly dilute SSBP/1. x axis is a logarithmic scale. The dashed red line represents 1,000 infected cells. Error bars show standard errors of the means. (b) Western blot of PrPC expression levels in Rov9 cells and the Rov9-B4 and -2A3 subclones. Amounts of 10 μg of total protein from induced (+ doxycycline) and uninduced (− doxycycline) cells were used. Lysates were PNGase treated to deglycosylate the PrPC so that the total amount of PrPC could be compared without the interference of glycosylated PrP. Molecular masses of size markers are shown on the left in kDa. (c) Immunofluorescent staining of PrPC in Rov9 and the two subclones Rov9-B4 and Rov9-2A3. Cells were either induced or uninduced for 48 h prior to staining for PrPC, which is colored green. Nuclei are falsely colored red.