Figure 5.
Analysis of PrPC interaction with ER chaperones. FRT cells were plated in 35-mm dishes in control (-), cholesterol depletion (Mev/βCD +), or sphingolipid depletion (FB1 +) conditions. The cells were then lysed in JS buffer and BiP, PDI, CNX, and CLT were immunoprecipitated (IP) in nondenaturing conditions with the specific antibodies (see Materials and Methods). The immunoprecipitates and 1/10 of the supernatants (Sup) were loaded on polyacrylamide gels. The gels were revealed for the presence of PrPC by Western blotting with the specific antibody (WB:α-PrP) by ECL.