NDNB1182 upregulates IFIT and endogenous retroviral element expression. (A) qRT-PCR and western blot of Ifit1 expression in MC38 treated with DMSO, Ganetespib, NDNB1182, or Hsp90α-i 12h, at the indicated concentrations for 6 hours. Cells were at 30% confluence when treated with the inhibitors. (B, C) Western blot of IFIT1, HSF1 and CDK4 in DU145 treated with DMSO or a concentration gradient of NDNB1182 for 12 hours. In (A, B), the quantification of band intensities was from one representative result of three repeated experiments that showed consistent results. (D–F) qRT-PCR result of Ifit and ERV elements in MC38, PPS and PyMT-7160 treated with DMSO, Ganetespib (1µM) or NDNB1182 (1µM) for 6 hours. In (D), MC38 confluence during the inhibitor treatment was indicated. (G) qRT-PCR result of Ifit1 expression in PPS treated with DMSO, NDNB1182 (1µM) and Palbociclib (2µM) for 6 hours. (H) qRT-PCR result of Ifnb1 expression in MC38 (50% confluence) treated with DMSO, Ganetespib (1µM) or NDNB1182 (1µM) for 6 hours. In (A, D–H), data represent mean ± SEM, N=3. #, P>0.05; *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, Student’s t-test.