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. 2015 Apr 7;16(4):518–527. doi: 10.1080/15384047.2015.1016661

Figure 5.

Figure 5.

TMZ-induced increase in DNA damage in hPMC2 downregulated MDA-MB-231 cells. MDA-MB-231 cells were transfected with hPMC2 miRNA and were either untreated or treated with 250 μM TMZ for 24 hours and analyzed via Western blot for APE1, γ-H2AX, and actin levels. (A) Representative protein gel blot of APE1 and actin levels of 2 independent experiments. (B) Representative western blot of γ-H2AX and actin levels of 2 independent experiments. (C) Quantification of γ-H2AX expression in MDA-MB-231 cells. γ-H2AX expression in control cells was given an arbitrary value of 1 and γ-H2AX expression in downregulated cells was plotted relative to control. Error bars indicate SEM of 2 independent experiments. a, significance (P < 0.05) vs. untreated cells; b, significance (P < 0.05) vs. control transfected cells with the same treatment. (D) Cells were analyzed using comet assay. Results are expressed in terms of the Olive Moment, which is the summation of each tail intensity integral value, multiplied by its relative distance from the center of the head and divided by the total comet intensity.