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. 2014 Aug 15;28(16):1800–1814. doi: 10.1101/gad.244368.114

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© 2014 Huang et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 2. — CDK9 is required for the proliferation of some HCC cell lines. (A) Competitive proliferation assay. G418-selected Venus⁺ cells were mixed with untransduced cells at 1:1 ratio and subsequently cultured in the presence of dox. The percentage of Venus⁺dsRed⁺ (shRNA-expressing) cells was determined at different time points (results at day 0 and day 14 are shown and are relative to day 0). Changes were used as readout of growth inhibitory effects. Values are mean + SD of three independent experiments. The graphs show the validation of the candidate shRNAs as well as control shRNAs (Ren.713, Myc.1891, and Rpa3.561) in MP1 murine HCC cells. (B) Immunoblots showing the knockdown induced by shRNAs expressed from TRMPV-neo in MP1 murine HCC cells. β-Actin was used as loading control. The numbers indicate protein levels relative to β-actin. (C) Competitive proliferation assay of control and candidate shRNAs expressed from TRMPV-neo in iMEFs, as described in A. Color code is as in A. (D) Immunoblots showing the knockdown induced by shRNAs expressed from TRMPV-neo in iMEFs. β-Actin was used as loading control. The numbers indicate protein levels relative to β-actin. (E,F) Competitive proliferation assay of control (Renilla and MYC) and CDK9 shRNAs expressed from TRMPV-neo-miR-E (E) or TRMPV-neo (F) in different murine (E) and human (F) cell lines, as described in A. The percentage of shRNA-expressing cells at day 14 relative to day 0 is shown. Values are mean + SD from two independent experiments.