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Figure 1


Fig. 1. Knockdown of both Dab2 and ARH results in accumulation of LDLR at the cell surface. (A) Western blot analysis of HeLa total cell lysates demonstrates that ARH, Dab2 and CHC were efficiently depleted following two rounds of transfection with short RNA duplexes specific for each target. As a control, MAPK levels remained constant in siRNA-treated cells. (B) Surface proteins were biotinylated and precipitated using streptavidin-linked agarose beads followed by western blot analysis for LDLR. Total cell lysates were also analyzed for LDLR on the same gel. (C) Densitometry was used to compare surface (biotinylated) LDLR to total cellular LDLR for each siRNA condition. The steady-state ratio of surface: total LDLR in control cells was set as 1. Depletion of ARH did not significantly change the amount of LDLR at the surface, whereas depletion of Dab2 caused a minor but significant increase (*P<0.01 as determined by t-test analysis). Depletion of both ARH and Dab2, or CHC, caused significant increases in relative surface LDLR (**P<0.001).





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