Deoxycholate, an Endogenous Cytotoxin/Genotoxin, Induces the Autophagic Stress-Survival Pathway: Implications for Colon Carcinogenesis
Figure 4
Immunohistochemical (a) and Western blot analysis (b), (c) of beclin-1
protein expression in NCM-460 cells in the absence and presence of 0.2 mM
DOC for 24 hours. (a) There is a dramatic increase in the staining of beclin-1 in
both the nucleus and cytoplasm of DOC-treated cells (right panel) compared with
control untreated cells (left panel). Beclin-1 staining also occurs in association
with the nucleolus where a ring of brown staining is observed in several cellular
profiles in DOC-treated cells. (100X oil objective; brown color of DAB indicates
positive beclin-1 staining; blue color indicates hematoxylin stain). (b) Western blot
indicating the increase in the beclin-1 61 kDa band after DOC treatment. (c) The
membranes were stained with Brilliant Blue G dye to confirm equal protein loading for western blot analysis. (d) Computerized densitometric evaluation of
bands shown in (b). ( indicates statistically significant (P < .05) differences in
mean Relative Densitometric Units (RDU) between treatment and untreated
control groups.)
