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. 2014 Feb 7;28(8):1636–1646. doi: 10.1038/leu.2014.19

Figure 6.

Figure 6

Inhibition of autophagy decreases primary CLL cell viability. (a) PBMCs from CLL patients (n=5) were incubated with varying concentrations of chloroquine or 3-MA for 48 h. Viability was determined by a CCK-8 colorimetric assay. Each sample was run in triplicate and was normalized to cells incubated without drug. Data represent the mean values±s.d. (b–d) PBMCs from CLL patients were transfected with autophagy gene-specific or nontargeting scrambled siRNA as indicated. Viability of CLL cells was assessed by flow cytometry following Annexin V-APC/PI staining. Results obtained with three different patient samples at (b) 24 h, (c) 48 h and (d) 72 h after transfection are shown. Viable cells (Annexin V-APC neg/PI neg cells) are represented and expressed relative to scrambled siRNA control, set at 100% viability. Expression levels of targeted genes were analyzed by real-time reverse transcription (RT)-PCR and were normalized to the 28S ribosomal RNA level of the same sample. RT-PCR values are expressed as relative fold change to the scrambled siRNA condition.