Figure 5.

CBB3001 Selectively inhibits the growth of F9 cells.

A. Actively growing F9 and NIH3T3 cells were treated with various concentrations of CBB3001 for 30 hours and examined. Cells were examined and cells images were acquired as in Figure 4A. Triplicated cells (technical repeats) were used for examination and one set of representative treated cells was shown.

B. Triplicated treated cells (technical repeats) from Figure 5A were harvested by trypsin digestion, diluted, and blindly spotted onto a hemacytometer. Cells in four corners of the hemacytometer were counted to obtain average cells per dish. The differences between control and CBB3001 cells in triplicated samples were plotted. Statistically significant differences were determined using a two-tailed equal-variance independent t-test. Different data sets were considered to be statistically significant when the P-value was <0.05 (*), 0.01 (**) or 0.001 (***).

C. The treated F9 cells were lysed with 0.5% SDS-containing lysis buffer and the levels of SOX2, OCT4, and actin (loading control) were monitored by Western blotting with specific antibodies indicated on the left, respectively, and quantified.