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. 2020 Jul 3;2(1):vdaa084. doi: 10.1093/noajnl/vdaa084

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© The Author(s) 2020. Published by Oxford University Press, the Society for Neuro-Oncology and the European Association of Neuro-Oncology.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Effects of methotrexate (MTX) treatment and leucovorin (LV) rescue were related to the degree of polyglutamylation in lymphoma cell lines. (A) Viability assays using cell lines after incubation with MTX (CellTiter-Glo). Each cell line was analyzed after 72 h of incubation. (b) Cells were treated with MTX for 24 h, followed by the addition of LV. Cell viability was assessed 48 h later. We defined EC₅₀ as the concentration of LV that recovered 50% cell viability. Data are shown as mean value ± SD from 3 independent experiments. *P < .01, compared with MTX. (C) Immunoblotting for folypolyglutamate synthetase (FPGS), γ-glutamyl hydrolase (GGH), and dihydrofolate reductase (DHFR) in the different cell lines. The internal control was α-tubulin. The relative expression of FPGS/GGH represents the ratio of FPGS/α-tubulin and GGH/α-tubulin calculated by densitometry. The FPGS/GGH ratio of HKBML is adjusted to 1. Data are shown as mean value ± SD from 3 independent experiments; *P < .05, **P < .01.