Figure 4

RHOXF2 is expressed in various cancer cell lines and modulates K562 resistance to DNA damaging agents. (a) In normal human tissues, RHOXF2 was exclusively detected in testis. Total RNA from 20 distinct tissues (mix of 3 different donors) was reverse transcribed into first strand cDNA and used as template for quantitative PCR. Three distinct pairs of primers covering exons 1, 2 and 4 were used. RHOXF2 expression level was calculated relative to the signal obtained for the amplification of housekeeping genes (CYCG, GUSB, ACTB, EEF1A1, GAPDH). (b) Representative example of immunohistochemical staining for RHOXF2 in normal testis. Section has been counterstained with Mayer’s hematoxylin. (c) RHOXF2 was detected in various tumor cell lines by western blotting. (d) RHOXF2 shRNA validation in the human chronic myelogenous leukemia cell line K562. WT, wild type. (e,f) Partial depletion of RHOXF2 increased K562 sensitivity to cisplatin and mitomycin C. Final numbers of viable cells were calculated in the absence of drug (inset). Student’s t-test; *P < 0.05; **P < 0.01; means ± standard error of the mean for n = 4 experiments.