Fig. 7: eEF2^T56 phosphorylation/expression is modulated and protein synthesis reduced, by mTORC1 inhibition of translation elongation in primary CLL cells.

A Protein lysates were prepared from primary CLL cells freshly isolated from the peripheral blood of consented patients (n = 18) and healthy B cells (HBC; n = 6) isolated from buffy coats. Western blots were performed and probed as indicated. Data are expressed as mean ± SEM, with p values were determined by two-tailed unpaired t-test (*p ≤ 0.05; ^**p ≤ 0.01). B MEC1 cells were treated with drugs for 24 h or (C) proliferating CLL cells are co-cultured for 5 days with NTL-CD40L/IL21 in the presence of drugs as indicated. Representative Western blots are shown. Blots shown are representative of n ≥ 3 individual biological replicates. D OPP assays were performed on drug-treated proliferating CLL cells to assess protein synthesis (n = 4). Data are expressed as mean ± SEM. p values were determined by one-way ANOVA, not corrected for multiple comparisons (*p ≤ 0.05, **p ≤ 0.001). E Graphs showing an average of the percentage of viable cells in treated primary CLL cells (n = 4). Data are expressed as mean ± SEM. p values were determined by an ordinary one-way ANOVA with Dunnett multiple comparisons test (*p ≤ 0.05).