Figure 7.

Effects of miR-200c on the distribution of c-Myc and β-catenin expressions in SKOV3 cell line. Cytoplasmic (C) and nuclear fractions (N) were analyzed by WB. β-actin and Lamin B1 are the housekeeping proteins used for cytoplasmic and nuclear extracts, respectively. (A) (i): empty vector (pCMV V.) and pCMV miR-200c SKOV3 cells treated with DMSO and 4 Gy irradiation (IR). (B) (i): vector and miR-200c transfected SKOV3 treated with 1.5 μM olaparib-only (OLAP) or in combination with 4 Gy irradiation. (C) (i): vector and miR-200c transfected SKOV3 treated with 5 μM olaparib-only (OLAP) treated vector and miR-200c or in combination with 4 Gy irradiation. The histograms reported below the WBs represent densitometric analysis by ImageJ of c-Myc and β-catenin, for each treatment. (ii,iii): the graphs indicate cytoplasmic and nuclear fractions of c-Myc respectively normalized to β-actin and Lamin B1 (iv,v): β-catenin expression in cytoplasmic and nuclear fractions. All the samples were normalized to the vector control SKOV3. The number of pixels from each protein signal imprinted on a film was normalized to the number of pixels of the respective housekeeping gene (β-actin or Lamin B1), calculated as a ratio. Three WB repetitions were performed with the same lysates and with protein lysates derived from three treatments upon olaparib and irradiation. Dunett’s multiple comparison statistical analysis was carried out with Prism 7 software, between pCMV V. DMSO/1.5μM OLAP/5 μM OLAP/IR-treated and the corresponding treated miR-200c transfected cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant.