Figure 4. HK2pep kills primary B‐CLL cells.

• A
  Representative IHCs in human reactive lymphoid tissue, where HK2 expression is mainly restricted to proliferating blasts within germinal centers (center) and in patient B‐CLL cells. Scale bar 100 μm.
• B–G
  Effect of cl‐HK2pep on human B‐CLL cells freshly isolated from patients. HK2‐expressing B‐CLL cells (B; GAPDH and respiratory complex III subunit UQCRC2 are cytosol and mitochondrial loading controls, respectively) undergo mitochondrial depolarization assessed by TMRM staining (C) and cell death, measured by cytofluorimetric analysis of Annexin V‐FITC and 7‐AAD staining (D‐G). Cells are treated with 5 μM cl‐HK2pep; PD150606 (50 μM) is pre‐incubated for 1 h. Experiments are carried out on samples from at least 10 patients and on CD19⁺, primary B lymphocytes from 5 healthy controls. In C, D, F, and G, data are presented as mean ± SEM; all data were obtained from three technical replicates in all analyzed patient samples. In (C, D) two‐way ANOVA: P < 0.0001 (cl‐HK2pep treatment versus cl‐HK2pep+PD150606 or cl‐SCRpep treatments); Bonferroni post‐test in graph ***P < 0.001 (cl‐HK2pep versus cl‐SCRpep and cl‐HK2pep versus PD150606+cl‐HK2pep), in F and G, Student's t‐test; ***P < 0.01, *P < 0.05.