Journal: PLOS One

Article Title: The EAAT1 aspartate/glutamate transporter is dispensable for acute myeloid leukemia cell growth and response to therapy

doi: 10.1371/journal.pone.0329048

Figure Lengend Snippet: A-C. Expression of SLC1A3 in AML cells across different patient genetic subgroups (A), according to FAB classification (B) and comparing newly diagnosed (ND) and relapsed (R) patients (C) in the BEAT-AML cohort. D-E. Expression of SLC1A3 in human AML cell lines (D) and of Slc1a1 , Slc1a2 and Slc1a3 in a mouse AML MLL-AF9 cell line (E) as quantified by qRT-PCR and expressed relative to β-actin ( ACTB ). F. Expression of SLC1A1 , SLC1A2 and SLC1A3 by human AML patient-derived xenograft cells isolated from mice treated with cytarabine or vehicle. Data obtained from (31). G. Expression of Slc1a1 , Slc1a2 and Slc1a3 by mouse AML cells (MA9−2 cell line) isolated from mice treated with induction chemotherapy (iCT; doxorubicin 3 mg/kg and cytarabine 100 mg/kg given in a 5 + 3 regimen) or vehicle. Data obtained from (19). Data are presented as mean ± SD. *P < 0.05; ***P < 0.001; ****P < 0.0001.

Article Snippet: U937, NB4, MV4–11, MOLM14, OCI-AML3, NOMO-1 and MONOMAC6 (MM6) human AML cell lines were obtained from the Leibniz Institute DSMZ and cultured at 37°C in a humidified incubator with ambient air and 5% CO 2 in RPMI1640 medium supplemented with 10% FBS, 100 I.U./mL penicillin-100 μg/mL streptomycin and 2 mM L-glutamine.

Techniques: Expressing, Quantitative RT-PCR, Derivative Assay, Isolation

Journal: PLOS One

Article Title: The EAAT1 aspartate/glutamate transporter is dispensable for acute myeloid leukemia cell growth and response to therapy

doi: 10.1371/journal.pone.0329048

Figure Lengend Snippet: A. Overall health of AML cells exposed to increasing concentrations of TFB-TBOA or UCPH-101 for 24 hours, as determined by the MTT assay. B. Half-maximal inhibitory concentration (IC50) for TFB-TBOA and UCPH-101 across 9 different human and mouse AML cell lines, based on the MTT assay. Each dot represents a different AML cell line. C. Viability of AML cells treated with TFB-TBOA (250 µM) or UCPH-101 (20 µM) for 24 hours as measured by flow cytometry. D. Cell cycle analysis of AML cells treated with UCPH-101 (20 µM) for 6 hours as measured by flow cytometry. Data are presented as mean ± SD. *P < 0.05; ***P < 0.001; ****P < 0.0001.

Article Snippet: U937, NB4, MV4–11, MOLM14, OCI-AML3, NOMO-1 and MONOMAC6 (MM6) human AML cell lines were obtained from the Leibniz Institute DSMZ and cultured at 37°C in a humidified incubator with ambient air and 5% CO 2 in RPMI1640 medium supplemented with 10% FBS, 100 I.U./mL penicillin-100 μg/mL streptomycin and 2 mM L-glutamine.

Techniques: MTT Assay, Concentration Assay, Flow Cytometry, Cell Cycle Assay

Journal: Genes & diseases

Article Title: RNA m 6 A reader YTHDF2 facilitates precursor miR-126 maturation to promote acute myeloid leukemia progression.

doi: 10.1016/j.gendis.2023.01.016

Figure Lengend Snippet: Figure 1 YTHDF2 is overexpressed in human AML and promotes AML cell growth/proliferation and colony-forming ability in vitro. (A) YTHDF2 expression in the healthy bone marrow (BM) and AML patients with different genetic backgrounds. AML MLL, MLL- rearranged AML. Data was obtained from MILE study and analyzed on Bloodspot (https://servers.binf.ku.dk/bloodspot/). t-test. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001. (B) Western blot of YTHDF2 levels in AML patients and cord blood. (Quantification was analyzed by ImageJ and showing the ratios of YTHDF2 to ACTB, which were normalized to the value in the BM-1 sample.) (CB), BM, or peripheral blood mononuclear cell (PBMC) from healthy donors. (C) Expression of YTHDF2 in AML patients with or without relapse/induction failure. Data was obtained from TARGET and analyzed on Xena (https://xenabrowser.net/). t-test. **, P < 0.01. (D) Western blot showing KD efficiency of YTHDF2 in AML cell lines (MM6 (MONOMAC6) and Molm13). (Quantification was analyzed by ImageJ and showing the ratios of YTHDF2 to GAPDH. All groups were normalized to shNS.) (E) Growth/proliferation changes of MM6 and Molm13 cells upon YTHDF2 KD as measured by MTT assay. Two-way Anova. ****, P < 0.0001. (F, G) Flow cytometry plots (F) and quantification (G) for apoptosis changes in MM6 and Molm13 cells upon YTHDF2 KD. (H, I) Numbers of colonies and repre- sentative pictures of colony-formation/replating assay (CFA) of MM6 and Molm13 cells with or without YTHDF2 KD. t-test. **, P < 0.01; ***, P < 0.001.

Article Snippet: MonoMac6 (MM6) was obtained from DSMZ and cultured in RPMI1640 (11875119, Thermo Fisher Scientific) supplemented with 10% FBS plus 2 mM L-glutamine (25030-081, Thermo Fisher Scientific), 1 non-essential amino acids (11140050, Gibco), 1 mM sodium pyruvate (11360070, Thermo Fisher Scientific), 10 mg/mL human insulin (12585014, Thermo Fisher Scientific).

Techniques: In Vitro, Expressing, Western Blot, MTT Assay, Flow Cytometry

Journal: Genes & diseases

Article Title: RNA m 6 A reader YTHDF2 facilitates precursor miR-126 maturation to promote acute myeloid leukemia progression.

doi: 10.1016/j.gendis.2023.01.016

Figure Lengend Snippet: Figure 4 YTHDF2 interacts with AGO2 and facilitates pre-mir-126 processing. (A) Overlapping of miRNA biogenesis regulators and potential YTHDF2 interacting partners identified by BioID of YTHDF2 N-terminus and C-terminus (data from a study published by J.Y. Youn et al).92 (B) Reciprocal co-IP between AGO2 and YTHDF2 using FLAG or control IgG antibodies in FLAG-tagged AGO2 or FLAG-tagged YTHDF2 overexpressing MM6 and THP1 cells. (C) Expression level changes of precursor and mature miR-126 after

Article Snippet: MonoMac6 (MM6) was obtained from DSMZ and cultured in RPMI1640 (11875119, Thermo Fisher Scientific) supplemented with 10% FBS plus 2 mM L-glutamine (25030-081, Thermo Fisher Scientific), 1 non-essential amino acids (11140050, Gibco), 1 mM sodium pyruvate (11360070, Thermo Fisher Scientific), 10 mg/mL human insulin (12585014, Thermo Fisher Scientific).

Techniques: Co-Immunoprecipitation Assay, Control, Expressing

Journal: Genes & diseases

Article Title: RNA m 6 A reader YTHDF2 facilitates precursor miR-126 maturation to promote acute myeloid leukemia progression.

doi: 10.1016/j.gendis.2023.01.016

Figure Lengend Snippet: Figure 5 MiR-126 OE partially rescues growth inhibition induced by YTHDF2 KD. (A) Levels of miR-126 on three time points (two, four, and seven days) after transfection of miR-126 mimics or negative control (NC) in MM6 cells. t-test. ****, P < 0.0001. (B) Growth changes of MM6 cells after miR-126 mimics transfection as measured by MTT. Two-way Anova. t-test. ***, P < 0.001. (C) Western blot of YTHDF2 in MM6 cells with or without YTHDF2 KD and/or miR-126 OE. (Quantification was analyzed by ImageJ and showing the ratios of YTHDF2 to GAPDH. All groups were normalized to shNS.) (D) Growth changes (as measured by MTT) of MM6 cells after YTHDF2 KD with transfection of miR-126 mimics or NC. Two-way Aanova. **, P < 0.01; ****, P < 0.0001. (E) Kaplan-Meier survival curves for BMT recipient mice transplanted with MA9 leukemic cells with or without miR-126 OE rescue after Ythdf2 cKO induced by poly I:C injection (n Z 8e10 each group). Log-rank test. NS Z not significant; *, P < 0.05.

Article Snippet: MonoMac6 (MM6) was obtained from DSMZ and cultured in RPMI1640 (11875119, Thermo Fisher Scientific) supplemented with 10% FBS plus 2 mM L-glutamine (25030-081, Thermo Fisher Scientific), 1 non-essential amino acids (11140050, Gibco), 1 mM sodium pyruvate (11360070, Thermo Fisher Scientific), 10 mg/mL human insulin (12585014, Thermo Fisher Scientific).

Techniques: Inhibition, Transfection, Negative Control, Western Blot, Injection

Journal: Genes & diseases

Article Title: RNA m 6 A reader YTHDF2 facilitates precursor miR-126 maturation to promote acute myeloid leukemia progression.

doi: 10.1016/j.gendis.2023.01.016

Figure Lengend Snippet: Figure 6 Downstream targets regulated by the YTHDF2/miR-126 axis in human AML. (A) QPCR showing expression changes of representative downstream target genes of miR-126 36 in MM6 cells after miR-126 OE with or without YTHDF2 KD. t-test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (B) Correlation between expression of reported miR-126 downstream target genes (ADAM9, ILK, TOM1, FCGR1A, FCN1, and LRCH4) and expression of YTHDF2 in AML patients. Data acquired from TCGA and analyzed at Xena. r, Pearson correlation coefficient.

Article Snippet: MonoMac6 (MM6) was obtained from DSMZ and cultured in RPMI1640 (11875119, Thermo Fisher Scientific) supplemented with 10% FBS plus 2 mM L-glutamine (25030-081, Thermo Fisher Scientific), 1 non-essential amino acids (11140050, Gibco), 1 mM sodium pyruvate (11360070, Thermo Fisher Scientific), 10 mg/mL human insulin (12585014, Thermo Fisher Scientific).

Techniques: Expressing