(A) Protein expression of CDH1 and ATG12–ATG5 in the
indicated pancreatic cancer cell lines transfected with control siRNA or
ATG5 siRNA. TUBB/β1-tubulin was used as a loading
control. For protein expression of CDH1 and ATG12–ATG5 in pancreatic
cancer cell lines with mutant KRAS, both short and long
exposures (respectively) are shown. KRAS mutation status is
indicated under the blots. (B) Fold change in mRNA levels of
CDH1, SNAI1, SNAI2,
TWIST1, ZEB1 and ZEB2 in
the indicated pancreatic cancer cell lines transfected with control siRNA or
ATG5 siRNA. GAPDH-normalized mRNA levels
in control cells were used to set the baseline value at unity. Data are mean
± s.d. n = 3 samples per group. * P
< 0.05. ** P < 0.01. *** P
< 0.001. (C) Immunofluorescence staining of CDH1 (green) in
HRas V12-expressing Atg7+/+
tumors or HRas V12-expressing
atg7-/- tumors. TO-PRO-3 (blue) was used to
stain nucleic acids. Scale bar: 20 μm.
Atg7+/+ or atg7-/- iBMK
cells transduced with HRas V12 were subcutaneously injected in
nude mice to form tumors. The graph shows the average relative intensity of CDH1
per cell evaluated using ImageJ, and data are mean ± s.d.
n = 4 random fields. *** P <
0.001.