Figure 3

HMGA1 silencing modulates the sensitivity to doxorubicin and LBH-589. (A) Western Blot analysis of FRO cells and FRO stably transfected with a short hairpin RNA targeting the HMGA1 gene (FRO shHMGA1) or with empty vector (FRO pLKO.1) using HMGA1 and β-actin antibodies. β-Actin was used as loading control (Left Panel). qRT-PCR analysis of HMGA1 mRNA in FRO pLKO.1 and FRO shHMGA1cells. Relative expression values indicate the relative change in HMGA1 mRNA expression levels between FRO pLKO.1 and FRO shHMGA1cells, normalized with β-actin, assuming that the mean value of FRO pLKO.1 was equal to 1. (Right Panel). The error bars represent the mean ± S.D. of three independent experiments performed in triplicate. (B) Cell viability assay of FRO pLKO.1 and FRO shHMGA1 cells treated with doxorubicin (Left Panel) or LBH-589 (Right Panel). Cells were exposed to the indicated doses of drugs and absorbance at 490 nm was measured after 72 hours. Values are the mean ± S.D. of three experiments performed in triplicate. Nonlinear regression lines were calculated using GraphPad Prism5. *p < 0.05 significance level compared with FRO pLKO.1. (C) IC50, inhibitory concentration that kills 50% of cell population and Resistance index calculated as IC50 FRO pLKO.1 /IC50 FRO shHMGA1 cell line.