Substitution of endogenous Reptin with D299N mutant leads to HuH7 cell growth arrest. Substitution of endogenous Reptin with D299N mutant leads to HuH7.

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Substitution of endogenous Reptin with D299N mutant leads to HuH7 cell growth arrest. Substitution of endogenous Reptin with D299N mutant leads to HuH7 cell growth arrest. Seven days after lentiviral transduction by siR2-resistant Flag-Reptin (FRrR2) or siR2-resistant Flag-Reptin D299N (FRD299NrR2), HuH7 cells were transfected with a control (siGL2) or an anti-Reptin siRNA (siR2). A, quantitative substitution of endogenous Reptin by Flag-Reptin or Flag-Reptin D299N was controlled by Western blot on whole-cell extracts 3 days after siRNA transfection. B, quantification of endogenous Reptin and Flag-Reptin expression. The graph shows the mean ± SD of 3 independent experiments. Total Reptin or Pontin amount was not significantly changed in any condition. C, adherent cells numbers were monitored at indicated time points with a Coulter counter. The graph shows the mean ± SD of 3 independent experiments (n = 3; ***, P < 0.001 for day 3 and day 6 by 2-way ANOVA followed by Bonferroni test). Note that SDs are not always visible because of their small size. Aude Grigoletto et al. Mol Cancer Res 2013;11:133-139 ©2013 by American Association for Cancer Research