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Figure 3: (a) Randomized alignment of control cells after 24 h; (b) randomized alignment of control cells after 48 h; (c) randomized alignment of control
cells after 72 h; parallel alignment of Ch-Np-treated cells [(d), (e), (f)]
Growth curve for control vs. treated CAF
Growth curve for untreated CAF showed a gradual
increase in the number of cells during 0-24 and 24-
48 h. After 48hr the number of cells almost doubled.
However, the 72-96 h time duration did not show a
doubling of cells. This experiment was repeated three
times and overall doubling time for untreated cells
was 25 ± 0.38 h [Figure 2]. In the case of treated cells,
during first 24 h, the cell count was less than the initially
seeded cells. Then, cells showed a gradual increase in
number. Importantly, the doubling rate of treated cells
was increased because the number of cells after 92 h
of culturing in the treated plate was less than that of
control cells, as shown in Figure 2. This experiment was
repeated three times and the overall doubling time for Figure 4: Expression of pluripotency markers in control and treated cells
treated cells was 30 ± 0.83 h [Figure 2].

Cellular morphology of CAF cells
Phase-contrast morphology of untreated and Ch-Np-
treated esophageal CAF was observed at 24, 48 and
72 h. Untreated CAF cells showed random growth
and cells were overlapping with each other, as shown
in Figure 3a, 3b, and 3c, whereas in the case of Ch-
Np-treated plates, the cells exhibited monolayers with
equal gaps and looked parallel to each other, as shown
in Figure 3d, 3e, and 3f. These cells did not overlap
with each other as was observed in the control CAF
cells. This seems to indicate that they changed their
malignant phenotype towards a normal phenotype by
Ch-Np treatment. Figure 5: Expression of differentiating markers in control and treated cells
Journal of Cancer Metastasis and Treatment ¦ Volume 2 ¦ July 29, 2016 ¦ 263