In vivo consequences of TNKS knockdown. (A) ED1 cells were infected with lentiviral shRNA constructs targeting TNKS1 or TNKS2 at two independent sites each, or the combination, and were selected with G418 (TNKS1 constructs and pLKO.1-CMV-Neo control), puromycin (TNKS2 constructs and TRC2 control), or the combination (dual control and dual shRNA). mRNA expression levels of the indicated species are detected by qPCR. (B) Axin 1 protein levels following stable TNKS knockdown are shown by immunoblot analysis. (C) Consequences on proliferation of stable TNKS knockdown in ED1 cells are shown in vitro as measured by luminescent cell viability assay 3 days after plating. (D) ED1 TNKS1 and TNKS2 shRNA dual transductants (or control) were injected into the flanks of athymic nude mice and tumor diameters measured twice weekly. Tumor growth rates are shown, N = 10 in each arm +/- SEM. (E) Time to the specified endpoint (designated as percent surviving) for the xenograft study is shown. Error bars represent SD of the mean. (* p ≤ 0.05).
Busch et al. BMC Cancer 2013 13:211 doi:10.1186/1471-2407-13-211