Fig. 3: PHGDH contributes to malignant B cell adaptation to ASNase treatment in vitro and in vivo. | Nature Communications

Fig. 3: PHGDH contributes to malignant B cell adaptation to ASNase treatment in vitro and in vivo.

From: Tumor metabolic adaptation induced by L-asparaginase reveals a vulnerability to PARP1/2 inhibitor in B-cell lymphomas

Fig. 3: PHGDH contributes to malignant B cell adaptation to ASNase treatment in vitro and in vivo.The alternative text for this image may have been generated using AI.

a Percentage of PHGDH activity in Eµ-Myc (#688) cells treated with DMSO or indicated concentrations of the PHGDH inhibitor BI-4916 for 24 h in Asn-containing medium (n = 3 independent experiments). b Percentage of dead Eµ-Myc (#688) cells (DAPI+) following 24 h of treatment with DMSO, ASNase (0.003 IU/ml) and/or BI-4916 (10 µM) in Asn-containing medium (n = 4 independent experiments). c Four-day proliferation of Eµ-Myc (#688) cells treated as in (b) (n = 5 independent experiments). d Total protein extracts prepared from Eµ-Myc (#506) cells stably expressing shRNA targeting the firefly luciferase (shSCR) or the murine Phgdh mRNA (two independent shPhgdh #1 and #2) were immunoblotted for indicated proteins. e Relative quantification of immunoblots presented in (d). (n = 4 independent experiments). f Percentage of dead Eµ-Myc (#506) cells (DAPI+) silenced (shPhgdh #1 or #2) or not (shSCR) for Phgdh mRNA following 24 h incubation in Asn-containing medium supplemented (+) or not (−) with ASNase (0.003 IU/ml) (n = 4 independent experiments). g Four-day proliferation of Eµ-Myc cells silenced or not (shSCR) for Phgdh mRNA, treated as in (f) (n = 5 independent experiments). h Survival curves of WT C57BL/6 mice intravenously injected with Eµ-Myc (#506) cells stably expressing shRNA control (shSCR) or targeting the murine Phgdh mRNA (shPhgdh #1), treated with Vehicle or ASNase every 48 h from day 7 until disease endpoint (n = 10 mice/group). i Total protein extracts prepared from Eµ-Myc cells isolated from BCL of C57BL/6 mice presented in h, were immunoblotted for the indicated proteins (shSCR-Vehicle, n = 3; shSCR-ASNase, n = 3; shPhgdh #1-Vehicle, n = 4; shPhgdh #1-ASNase, n = 4 mice). V Vehicle, A ASNase. The samples derive from the same experiments but different gels for PHGDH, PSAT1, ERK2, and another for ASNS and ERK2 were processed in parallel. j Relative quantification of PHGDH protein levels presented in (i) Data are normalized to the control condition shSCR-Vehicle. Data are expressed as mean ± SD (a, b, e, f, j) or ± SEM (c, g). P-values are from one-way Anova followed by Tukey’s test (a, e), 2-way Anova followed by Tukey’s t-test (b, f, j), t-test (c, g), log-rank test (h), and indicated as ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001. For detailed individual P-value, please refer to the Source data table.

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