Figure 6.

Stabilization of a Bag6-Associated MLP upon RNF126 Knockdown in Cells

(A) HEK293T cells expressing N3a-PrP were incubated without or with 10 μM MG132 for the indicated times and analyzed by immunoblotting for N3a-PrP. Two exposures are shown and the ubiquitinated species indicated.

(B) Lysates from cells expressing N3a-PrP (top panels) or wild-type PrP (bottom panels) were subjected to immunopurification using anti-Bag6 or control antibodies. Bag6, N3a-PrP, and PrP were followed during the purification by immunoblot. Two exposures of N3a-PrP are shown.

(C) HEK293T cells expressing N3a-PrP were transfected with the indicated siRNAs for 72 hr. The lysates were analyzed for ubiquitination activity toward Bag6-Luc (top), and for RNF126, N3a-PrP, and actin by immunoblot (bottom).

See also Figure S6.