Figure 1.
Impact of PTEN loss and PIK3CA gain on AKT activity and BCR prosurvival signaling. Loss of PTEN expression but not PIK3CA (over)expression is associated with hyperphosphorylation and overactivation of AKT. On the contrary, both loss of PTEN and, to a lesser extent, gain of PIK3CA decrease dependance on the prosurvival signaling from BCR. (A) Western blot. Confirmation of the upregulation of PIK3CA in the 4 PIK3CA UP cell lines MINO, Z138, UPF1H, and UPF19U and the loss of PTEN in the PTEN KO clones MINO and UPF1H. Increase of phospho-AKT (ser 473) expression in both PTEN KO cell lines MINO and UPF1H (for each sample, n = 2). (B) AKT activity as measured using genetically encoded FRET-based biosensor. Significantly increased AKT kinase activity in all PTEN KO cell lines compared with respective control cell lines; technical triplicates; (C) FRET assay. Variable effect of PIK3CA (over)expression on AKT activity; technical triplicates. (D) PTEN loss increases survival of MCL cells with knockout of BCR gene (n = 3). (E) Transgenic (over)expression of PIK3CA in MINO and JEKO-1 but not in UPF1H increases survival of MCL cells with knockout of BCR gene (n = 3). (F) AKT activity as measured using genetically encoded FRET-based biosensor. AKT activity in PTEN KO cell lines is higher than that of respective cell lines. In addition, after exposure to BTKi ibrutinib for 3 hours, AKT activity in PTEN KO cell lines remains higher than AKT activity in the respective control cell lines (n = 3). (G) Proliferation assay implemented 72 hours after exposure to ibrutinib (0.1 and 1 μM); the cellular proliferation of the treated cells was normalized to the cellular proliferation of the untreated cells (n = 3); data are represented as mean ± standard deviation (SD); ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001; (N) represents the number of biological replicates. CTRL, controls; DMSO, dimethyl sulfoxide; ns, not significant.

Impact of PTEN loss and PIK3CA gain on AKT activity and BCR prosurvival signaling. Loss of PTEN expression but not PIK3CA (over)expression is associated with hyperphosphorylation and overactivation of AKT. On the contrary, both loss of PTEN and, to a lesser extent, gain of PIK3CA decrease dependance on the prosurvival signaling from BCR. (A) Western blot. Confirmation of the upregulation of PIK3CA in the 4 PIK3CA UP cell lines MINO, Z138, UPF1H, and UPF19U and the loss of PTEN in the PTEN KO clones MINO and UPF1H. Increase of phospho-AKT (ser 473) expression in both PTEN KO cell lines MINO and UPF1H (for each sample, n = 2). (B) AKT activity as measured using genetically encoded FRET-based biosensor. Significantly increased AKT kinase activity in all PTEN KO cell lines compared with respective control cell lines; technical triplicates; (C) FRET assay. Variable effect of PIK3CA (over)expression on AKT activity; technical triplicates. (D) PTEN loss increases survival of MCL cells with knockout of BCR gene (n = 3). (E) Transgenic (over)expression of PIK3CA in MINO and JEKO-1 but not in UPF1H increases survival of MCL cells with knockout of BCR gene (n = 3). (F) AKT activity as measured using genetically encoded FRET-based biosensor. AKT activity in PTEN KO cell lines is higher than that of respective cell lines. In addition, after exposure to BTKi ibrutinib for 3 hours, AKT activity in PTEN KO cell lines remains higher than AKT activity in the respective control cell lines (n = 3). (G) Proliferation assay implemented 72 hours after exposure to ibrutinib (0.1 and 1 μM); the cellular proliferation of the treated cells was normalized to the cellular proliferation of the untreated cells (n = 3); data are represented as mean ± standard deviation (SD); ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001; (N) represents the number of biological replicates. CTRL, controls; DMSO, dimethyl sulfoxide; ns, not significant.

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