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. Author manuscript; available in PMC: 2015 May 1.
Published in final edited form as: Mitochondrion. 2013 Oct 29;16:55–64. doi: 10.1016/j.mito.2013.10.004

Figure 3.

Figure 3

The apoptotic effect of NaBt on human CRC cells. SW480 and HCT116 cells were treated with 1, 2.5 and 5 mM NaBt for 12 and 24 h. (A-B) At the end of treatments, total cells were collected and stained with annexin V/PI and analyzed for apoptotic cell population as mentioned in ‘Materials and Methods’. Data are mean ± SD of triplicate samples in each case. *, P<0.05, significantly different from control by one-way ANOVA followed by Dunnett’s test. (C) In similar treatments as detailed above, total cell lysates were prepared as described in ‘Materials and Methods’. SDS-PAGE and western blot analysis were performed for Bcl-2 and total as well as cleaved PARP. Membranes were stripped and re-probed with anti-beta-actin antibody to ensure equal protein loading. (D) In similar treatment, semiquantitative RT-PCR was done for mRNA levels of Bcl-2 and GAPDH in both cell lines as detailed in ‘Materials and Methods’. (E) Effect of NaBt treatment on cleavage of caspase-3 and its total level were analyzed in SW480 and HCT116 at 24 h using specific antibody by SDS-PAGE and western blot analysis and membrane was stripped and re-probed with anti-beta-actin. Numbers on top of the bands represent fold changes in band intensity as compared to control as determined by densitometric analysis of the bands and corrected for beta-actin or GAPDH loading control for western blot or PCR band, respectively (C-E).