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. Author manuscript; available in PMC: 2016 Aug 18.
Published in final edited form as: J Cancer Ther. 2014 Oct 28;5(12):1092–1113. doi: 10.4236/jct.2014.512114

Figure 4.

Figure 4

Knockdown of EWS expression inhibited cell proliferation and induced differentiation with modulation of expression of specific molecules in human Ewing’s sarcoma SK-N-MC and RD-ES cell lines. (A) In situ methylene blue staining to assess decrease in cell proliferation and increase in morphological features of neuronal differentiation. (B) Measurement of dimensions (cell width, cell length, and extension length) of the differentiated cells. Transfection of cells with scrambled shRNA plasmid (0.5 mg/ml) or EWS shRNA plasmid (0.5 mg/ml) for 72 h. Mean values of three independent experiments were shown. Significant difference between EWS shRNA and scrambled shRNA was indicated by *P < 0.05 or **P < 0.01. (C) Induction of differentiation with modulation of expression of specific molecules. Treatments: scrambled shRNA plasmid (0.5 μg/ml) transfection for 72 h, EWS shRNA plasmid (0.5 μg/ml) transfection for 72 h, 100 μM TFL treatment for 24 h, and EWS shRNA palsmid (0.5 μg/ml) transfection for 48 h + 100 μM TFL treatment for last 24 h. Protein samples were used for Western blotting to examine changes in expression of specific molecules.