FIG. 5.

Continuous HRV2 internalization leads to degradation of mutant but not wt LDLR. RF3 and ΔYC cells were preincubated in serum-free Ham F-12 medium, and HRV2 at 1,500 TCID₅₀/cell was internalized for 6 h. Cells were cooled, washed, and processed for indirect immunofluorescence microscopy for the detection of LDLR (green) and LAMP2 (red). Nuclei were stained with Hoechst dye (for epifluorescence) and DRAQE5 (for confocal microscopy). (A) Conventional epifluorescence microscopy. All images were taken with the same exposure time in the respective channel and identical settings were used for illustration with the Axiovision software. Overlay images are shown. Arrowheads indicate perinuclear and arrows indicate plasma membrane localization of the receptors. (B) Confocal images were taken by using the same laser power and exposure time in the respective channel. Multicolor images shown were obtained with identical gray level settings in each channel. Of 20 sections through the cells, the focal plane through the nucleus is depicted. Bar, 2 μm.