Figure 5.
Impact of CSF on the expression of methotrexate target proteins in leukemia cells. (A) Immunoblots showing the effects of CSF on methotrexate target proteins. Leukemia cell lines were cultured in regular media or CSF for 48 hours. Protein lysates were then collected for immunoblotting with DHFR, TYMS, ATIC, or β-actin antibodies. (B-D) Dose-response curves for wild-type (WT) DHFR or mutant DHFR (mDHFR; L22F, F31S) NALM-6 (B), REH (C), and Jurkat (D) leukemia cells treated with different concentrations of the methotrexate. Leukemia cell viability was assessed after 48 hours of drug treatment using the CellTiter-Glo luminescent cell viability assay. Error bars represent the mean ± SD of 3 technical replicates. LogIC50 and bottom values with confidence intervals were calculated from the dose-response curves and are shown in the supplemental Table.

Impact of CSF on the expression of methotrexate target proteins in leukemia cells. (A) Immunoblots showing the effects of CSF on methotrexate target proteins. Leukemia cell lines were cultured in regular media or CSF for 48 hours. Protein lysates were then collected for immunoblotting with DHFR, TYMS, ATIC, or β-actin antibodies. (B-D) Dose-response curves for wild-type (WT) DHFR or mutant DHFR (mDHFR; L22F, F31S) NALM-6 (B), REH (C), and Jurkat (D) leukemia cells treated with different concentrations of the methotrexate. Leukemia cell viability was assessed after 48 hours of drug treatment using the CellTiter-Glo luminescent cell viability assay. Error bars represent the mean ± SD of 3 technical replicates. LogIC50 and bottom values with confidence intervals were calculated from the dose-response curves and are shown in the supplemental Table.

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