Figure 4. Akirin-2 mediated chemoresistance could be abolished by RNAi technology as measured by (a) cleaved (c)Caspase-3/-7 activity assay, (b) cCaspase-3 and -7 and (c) cleaved (c)poly(ADP-ribose) polymerase-1 (cPARP-1) Western Blots.

a. In relation to mock transfected T98G cells Akirin-2 knock down (kd) was able to induce cell death to significantly greater extents in dimethylsulfoxide (DMSO) as well as in TMZ (400 μg/ml, 24 h) treated samples (n = 6; triple values; boxplot: bold line = median; box = upper and lower quartile; whisker = 1.5-fold of interquartile range, circles = outlier). b. Results were confirmed by Western Blot using specific antibodies directed against cCaspase-3 and -7, respectively. c. PARP-1 Western Blot using two different antibodies specifically directed against cleaved PARP-1 (bottom) or both uncleaved and cleaved PARP-1 (top) additionally approved results. Equal protein loading was confirmed by detection of glycerinaldehyde-3-phosphate-dehydrogenase (GAPDH) or heat shock protein 90 (Hsp90), and efficiency of knock down was proven by qRT-PCR for all experiments in parallel. Representative examples of two independent experiments are shown.