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. 2022 Jan 13;107(8):1796–1814. doi: 10.3324/haematol.2021.279924

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Copyright© 2022 Ferrata Storti Foundation

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Figure 7. — PTEN expression is downregulated in human chronic lymphocytic leukemia B cells. (A and B) Flow cytometric analysis of PTEN expression (B) in human blood samples from healthy donors (HD) B1 cells (black), B2 cells (green) and mutated chronic lymphocytic leukemia (M-CLL, blue) and unmutated CLL (U-CLL, red) patients. The histograms indicate the fluorescence intensity of PTEN protein expression of representative samples after gating on CD19⁺ and CD5⁺ cells from the U-/M-CLL patients and HD B1 cells (A). For the HD B2 cells, we gated on CD19⁺ and CD5^- cells (A, left). (C) Representative isotype control (C, left) and size control (C, right) of the flow cytometric analysis of PTEN expression in human blood samples from HD B1 cells (black), B2 cells (green) and M-CLL (blue) and U-CLL (red) patients. (D) Quantification of human blood samples from HD (age 60+) and CLL patients with and without a mutation in the IGHV gene (U-CLL and M-CLL). The plot indicates the mean fluorescence intensity (MFI) of PTEN protein expression after gating on CD19⁺ and CD5⁺ cells for the M-/U-CLL patients and the HD B1 cells. For the HD B2 cells we gated on CD19⁺ and CD5^- cells. The bars include median and standard deviation. P-values were obtained using two-tailed Student's t-test (*P<0.05; **P<0.01; ***P<0.001). Every dot or triangle represents an individual. (E) Real-time quantitative polymerase chain reaction (qRT-PCR) analysis of human blood samples from HD and U-/M-CLL. The plot indicates the levels of PTEN mRNA expression relative to the expression of the housekeeping gene GAPDH, including median and standard deviation. P-values were obtained using two-tailed Student's t-test (**P<0.01; ****P<0.0001). Every dot or triangle represents an individual. (F) Relative PTEN mRNA and protein expression in B cells from U-/M-CLL patients compared to the averaged expression of HD control. Red: samples show high PTEN mRNA/protein expression relative to HD, black: PTEN expression is similar relative to HD, grey: CLL patients have lower PTEN/mRNA levels relative to HD.