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. 2009 Apr 24;284(17):11706–11716. doi: 10.1074/jbc.M805990200

FIGURE 8.

FIGURE 8.

Reduction of synphilin-1A expression decreases the steady-state levels of SIAH-1 and its substrates synphilin-1 and DCC. A, SH-SY5Y cells were transfected with HA-synphilin-1A, in the presence of siRNA control or siRNA to synphilin-1A (Sph-1A). HA-synphilin-1A from total cell lysates was detected by Western blot using anti-HA antibody (upper panel). B, SH-SY5Y cells were transfected with siRNA control or siRNA to synphilin-1A. The endogenous levels of SIAH-1 from total cell lysates were detected by Western blot using the 990 anti-SIAH-1 antibody (8) (upper panel). Middle panel shows the levels of endogenous synphilin-1A with anti-synphilin-1A antibody. C, SH-SY5Y cells were transfected with HA-synphilin-1, in the presence of siRNA control or siRNA to synphilin-1A. HA-synphilin-1 from total cell lysates was detected by Western blot using anti-HA antibody (upper panel). Middle panel shows the levels of endogenous SIAH-1 with 990 anti-SIAH-1 antibody. D, SH-SY5Y cells were transfected with HA-DCC, in the presence of siRNA control or siRNA to synphilin-1A. HA-DCC from total cell lysates was detected by Western blot using anti-HA antibody (upper panel). Middle panel shows the levels of endogenous SIAH-1 with 990 anti-SIAH-1 antibody. Lower panels in A-D show the loading control of protein extracts determined by incubating the membranes with anti-actin antibody.