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. 2018 Sep 11;9(9):930. doi: 10.1038/s41419-018-0952-8

Fig. 3. TAp73 downregulation sensitizes melanoma cells towards cisplatin and carboplatin treatment.

Fig. 3

a Viability analysis was performed in S melanoma cell lines after the transfection of siRNA against TAp73 (si1 TAp73) or non-silencing control siRNA (siCtr.) and the additional treatment with up to 20 µM cisplatin (CDDP) for 72 h. The assay signals were normalized to the appropriate untreated cell signals. Exemplary data of one experiment with five replicates are shown. (Mean values ± SD). b Apoptotic cell fraction (in %) was analysed after the treatment of siRNA-transfected melanoma cell lines (si1TAp73, si2TAp73, siCtr.) with 2.5 µM cisplatin (CDDP) for 72 h via cell cycle analysis. Transfected and untreated cell samples were used for the detection of basal apoptotic cell fractions (Ctr.). Exemplary data of one experiment with triplicates are shown. (Mean values ± SD). c Viability analysis was performed on R melanoma cells transfected with HA-TAp73α plasmid (TAp73α) or pcDNA3 control-plasmid (Ctr.), with or without treatment with cisplatin (CDDP, up to 20 µM) for 72 h. Assay signals were normalized to untreated cell signals. Exemplary data of one experiment with five replicates are shown. (Mean values ± SD). d Comparison of cisplatin EC50 values (blue bar) and TA TP73 expression (grey bar) in metastatic melanoma cell lines. TAp73 mRNA expression (TA TP73) was normalized to Actin expression (Mean values ± SD; n = 3). EC50 values were analysed using the data of the appropriate viability assays (n = 3) after cisplatin treatment (up to 20 µM) for 72 h. e Representative pictures of pH2AX (in red) and YO-PRO (in green) staining in S melanoma cells transfected with siRNA against TAp73 (si1 TAp73) or control siRNA (siCtr.) with or without additional cisplatin treatment (CDDP, 24 h, 5 µM). The white bar represents 25 µm. Right graph: pH2AX density (foci/cell) after cisplatin treatment was normalized to the basal pH2AX density in the appropriate cells without treatment set as 1 (Mean values ± SD, n = 3)