EZH2 expression is reduced at post-transcriptional level but is not involved in MM cells response to DZNep treatment.

<p>(<b>a</b>) MMCLs were treated with either vehicle (0) or indicated concentrations of DZNep for 72 h. Western blots were performed with indicated antibodies. Anti-GAPDH Ab was used for loading and transfer control. The experiment has been repeated three times. (<b>b</b>) Responsive 8226 and resistant LP1 cell lines were treated for different time intervals (<b>upper part</b>) or 72 h (<b>lower part</b>). The transcriptional expression of <i>EZH2</i> (<b>upper part</b>) or <i>BMI1</i>, <i>EED</i> and <i>SUZ12</i> (<b>lower part</b>) was studied by qRT-PCR using the ΔΔC_t method with <i>RPLP0</i> as internal standard. The fold change was calculated as 2^{−ΔΔ<i>Ct</i>}. Indicated values corresponded to the mean ± SD from at least three independent experiments. (<b>c</b>) The responsive 8226 and JJN3 and the resistant LP1 cells were treated with vehicle, MG-132 (100 nM for 24 h), DZNep (1 µM for 24 h) or both. Cell were harvested; total proteins were purified, separated by SDS-PAGE and analyzed by Western blot with the indicated Abs. (<b>d</b>) 8226 cells were treated with CHX (100 µM for 1 h) then with vehicle or DZNep (1 µM for 24 h). Total proteins were purified, separated by SDS-PAGE and analyzed by Western blot with the indicated Abs.