Biology Reference
In-Depth Information
Figure 7.
Endocytosis of PrP increases ferritin iron content. (A) PrP
C
-cells exposed to 1 μg/ml of 3F4
for 5 days were radiolabeled with
59
FeCl
3
for 4 hr, and lysates were fractionated on a non-denaturing
gel and auto-radiographed (lanes 1 and 2). Equal aliquots of the same samples were boiled in SDS-
containing sample buffer and fractionated in duplicate by SDS-PAGE followed by immunoblotting
with PrP specific antibodies 3F4 and 8H4 (lanes 3-6). Subsequently, the membranes were re-probed
for ferritin, Tf, TfR, and
-actin (lanes 7 and 8). (B) Quantification by densitometry shows an increase
in ferritin iron and TfR levels, and insignificant change in Tf levels in 3F4 exposed cells. Values are
mean±SEM of three independent experiments. *p<0.001 compared to untreated cells. (C) Estimation
of LIP after exposing the cells to 1 μg/ml of 3F4 or anti-Thy-1 antibody for 5 days shows insignificant
difference between untreated and 3F4 treated PrP
C
cells, and a decrease in anti-Thy-1 treated cells.
*p<0.001. n = 5.
β
