Additional file 2: Figure S1. of Aurora kinase targeting in lung cancer reduces KRAS-induced transformation

KRAS regulates AURKA and AURKB mRNA levels. a Expression of KRAS, AURKA, and AURKB was analyzed by real-time quantitative PCR in immortalized primary epithelial lung cells (SALEB) and their isogenic KRAS-transformed counterpart (SAKRAS), showing that KRAS mRNA expression positively correlates with both AURKA and AURKB mRNA expression. Statistical significance was measured by Student’s t-test (*p < 0.05) and error bars represent average ± 1s.d. b mRNA expression of KRAS, AURKA, and AURKB was analyzed by real-time quantitative PCR in H1703 lung cancer cells engineered to express KRASG12V inducibly (G12V) in comparison to empty vector-transfected H1703 cells (TrexB). To induce KRAS expression, cells were treated with 2 μg/mL doxycycline for the indicated times. Statistical significance was measured by Student’s t-test (*p < 0.05) by comparing treated samples (DOX) to untreated control samples (MOCK). Error bars represent average ± 1s.d. c Fluorescence microscopy images of A549 and H358 stable cells with inducible expression of 2 different shRNAs targeting KRAS (shKR#1 and shKR#2), AURKA (shAKA#1 and shAKA#2), AURKB (shAKB#1 and shAKB#2) or a non-targeting shRNA (shCtrl) showing induction of red fluorescent reporter protein (RFP) expression upon treatment with 2 μg/mL doxycycline for 5 days (DOX) when compared to untreated (MOCK) cells. Brightfield images of the same fields are included for comparison. d mRNA expression of KRAS, AURKA, and AURKB was analyzed by real-time quantitative PCR in A549 and H358 stable cells with inducible expression of 2 different shRNAs targeting KRAS (shKR#1 and shKR#2) or a non-targeting shRNA (shCtrl). Cells were either treated with 2 μg/mL doxycycline (DOX) for 5 days to induce shRNA expression or left untreated (MOCK). Statistical significance was measured by Student’s t-test (*p < 0.05, **p < 0.01) by comparing treated samples (DOX) to untreated control samples (MOCK). Error bars represent average ± 1s.d. (PDF 648 kb)