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.