Research Article
YiQiFuMai Powder Injection Protects against Ischemic Stroke via Inhibiting Neuronal Apoptosis and PKCδ/Drp1-Mediated Excessive Mitochondrial Fission
Figure 6
YQFM regulates mitochondrial fission through inhibition of PKCδ-mediated Drp1 activation. Neurons were pretreated with YQFM (400 μg/ml), Rottlerin (1 μM) or Mdivi-1 (25 μM) for 6 h and then incubated with H2O2 (100 μM) for 6 h. (a) Mitochondrial morphology stained with MitoTracker Deep Red FM. Bar = 5 μm. (b) Protein expression of Drp1 and p-Drp1 (Ser616). Expression of Drp1 and PKCδ in the mitochondrial (c) or cytosolic (d) fraction was determined using Western blot. Coimmunoprecipitation of Drp1 and PKCδ in the mitochondrial (e) or cytosolic (f) fraction was detected by Western blot with indicated antibodies. Cytosolic or mitochondrial fractions were subjected to coimmunoprecipitation with anti-PKCδ antibody or anti-Drp1 antibody, and then the precipitates were analyzed by immunoblotting with anti-PKCδ and anti-Drp1 antibodies. IgG was loaded as a negative control. IB: immunoblotting, IP: immunoprecipitation and IgG: immunoglobulin G. versus control, versus H2O2, versus H2O2.
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