Journal: Stem Cell Research & Therapy

Article Title: Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro

doi: 10.1186/s13287-020-01671-1

Figure Lengend Snippet: Results of RT-PCR production of spermatogonial stem cells and EL4 cells. The expression of PLZF, GFRα-1, and Integα-6 in spermatogonial stem cells and H2Kb for EL4 cells. β-actin was included as a housekeeping gene

Article Snippet: Finally, in the present study, three main groups were designed, and by accessing the culture collection of Pasteur Institute, Tehran, Iran, the mouse acute lymphoblastic leukemia cell line EL4 was prepared.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro

doi: 10.1186/s13287-020-01671-1

Figure Lengend Snippet: Results of flow cytometry in the mixture of cancer cells and spermatogonial stem cells after microfluidic isolation. a The results obtained from cells without adding antibodies. b The results of EL4 cell composition and SSCs (in vitro tumor model). c The results of cell percentage after passing microfluidic of outer outlet. d The results of the percentage of cells associated with the inner outlet. Q1, the range of PLZF positive cells and negative H2k cells; Q2, the range of PLZF positive cells and positive H2k cells, Q3, the range of negative PLZF cells and negative H2k cells; Q4, the range for negative PLZF and positive H2k

Article Snippet: Finally, in the present study, three main groups were designed, and by accessing the culture collection of Pasteur Institute, Tehran, Iran, the mouse acute lymphoblastic leukemia cell line EL4 was prepared.

Techniques: Flow Cytometry, Isolation, In Vitro

Journal: Stem Cell Research & Therapy

Article Title: Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro

doi: 10.1186/s13287-020-01671-1

Figure Lengend Snippet: SSC colonies of mouse neonate spermatogonial stem cell after 2 weeks of culture in free-growth factors DMEM/F12, 1 week after primary culture, and EL4 tumor cell. a Complete colony of spermatogonium cells. b Tumor cells. Scale bars = 100 μm

Article Snippet: Finally, in the present study, three main groups were designed, and by accessing the culture collection of Pasteur Institute, Tehran, Iran, the mouse acute lymphoblastic leukemia cell line EL4 was prepared.

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro

doi: 10.1186/s13287-020-01671-1

Figure Lengend Snippet: Part A is for the PLZF marker for SSC cells and part B is for tumor cell CD45 markers and the MERGE-shaped image of the above images and DAPI staining. Based on flow cytometry, in the first row, the control group, which contains the SSC and EL4 cell composition, is seen, and both types of cells are seen. In the 2nd row is the output from the outer inlet, which indicates a higher number of EL4 cell. The 3rd row refers to the output of the inner outlet which indicates a higher number of PLZF markers, the SSC cell

Article Snippet: Finally, in the present study, three main groups were designed, and by accessing the culture collection of Pasteur Institute, Tehran, Iran, the mouse acute lymphoblastic leukemia cell line EL4 was prepared.

Techniques: Marker, Staining, Flow Cytometry, Control

Journal: Immunity

Article Title: A Thpok-directed transcriptional circuitry promotes Bcl6 and Maf to orchestrate T follicular helper differentiation.

doi: 10.1016/j.immuni.2019.06.023

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Transient transfection into RLM-11 cells was performed using Amaxa nucleofection Cell Line Nucleofector (solution L and EL4 program).

Techniques: Generated, Recombinant, Purification, Staining, Plasmid Preparation, Reporter Assay, Expressing, Transduction, Transgenic Assay, Software

Journal: PLoS ONE

Article Title: Transcription Factors Oct-1 and GATA-3 Cooperatively Regulate Th2 Cytokine Gene Expression via the RHS5 within the Th2 Locus Control Region

doi: 10.1371/journal.pone.0148576

Figure Lengend Snippet: (A) Naïve CD4 T cells were activated, transduced with Oct1-MIEG3, Oct2-MIEG3, or control (empty) MIEG3 retroviral vector, and differentiated into Th2 cells. Cytokine expression was measured by intracellular cytokine staining. The plots show the percentages of differentiated cells expressing a particular cytokine. Data are representative of three independent experiments with similar results. (B) The results in (A) were shown as bar graphs. Statistical differences between groups were analyzed by Student’s t -test (*: P < 0.05). (C) Expression levels of Th2 cytokine mRNA. Activated T cells were transduced with Oct-1-MIEG3 or control MIEG3 vector as in (A). GFP+ cells were sorted from samples used in (A) and re-stimulated with anti-CD3 antibody for 24 h. The mRNA expression levels for selected Th2 cytokines were measured by quantitative PCR. Statistical differences between groups were analyzed by Student’s t -test (*: P < 0.05). (D) The effect of Oct1 siRNA on the expression of the Il4 gene. EL4 cells were transfected with control or Oct1 siRNA. The mRNA expression levels for the Il4 gene was measured by quantitative RT-PCR. Bars indicate mean ± SD (n = 3). Statistical difference between groups was analyzed by Student’s t -test (*: P < 0.05). (E) Effect of Oct-1 overexpression on cytokine expression in Th1 cells. Naïve CD4 T cells were transduced with Oct1-MIEG3 or control MIEG3 retroviral vector as in (A) and differentiated into Th1 cells. Cytokine expression was measured by intracellular cytokine staining. The plots show the percentages of cells expressing a particular cytokine. Data are representative of three independent experiments with similar results. (F) IFN-γ-positive cells in (D) were shown as a bar graph.

Article Snippet: The T lymphocyte EL4 cell line and the Phoenix-Eco cell line were purchased from the Korean Cell Line Bank (Laboratory of Cell Biology, Cancer Research Center and Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea).

Techniques: Transduction, Control, Retroviral, Plasmid Preparation, Expressing, Staining, Real-time Polymerase Chain Reaction, Transfection, Quantitative RT-PCR, Over Expression

Journal: PLoS ONE

Article Title: Transcription Factors Oct-1 and GATA-3 Cooperatively Regulate Th2 Cytokine Gene Expression via the RHS5 within the Th2 Locus Control Region

doi: 10.1371/journal.pone.0148576

Figure Lengend Snippet: (A) EL4 cells were transfected with expression vectors (Oct1-CMV, Oct2-CMV, GATA-3-CMV, or control CMV) and the RHS5-IL4 reporter construct by electroporation. Cells were allowed to rest for 18 h, followed by treatment with PMA (50 ng/ml) and ionomycin (1 μM) for 4 h. Luciferase activity was then measured. Data are representative of three independent experiments with similar results (NT = no treatment). (B) Mutation sites of Oct-1-binding sites within Il4 promoter and RHS5. Oct-1-binding sites, close to GATA-3-binding site, within Il4 promoter were deleted as indicated (upper), or Oct-1 binding site adjacent to GATA3 binding site within RHS5 was mutated as indicated (lower). (C) RHS5(WT)-IL4P(WT) reporter construct or RHS5(mtOct1)-IL4P(WT) construct (RHS5-IL4P reporter construct bearing mutation at Oct-1 binding site within RHS5) were transfected into EL4 cells with expression vectors (Oct1-CMV, GATA-3-CMV, or control CMV) by electroporation. Cells were allowed to rest for 18 h, followed by treatment with PMA (50 ng/ml) and ionomycin (1μM) for 4 h. Luciferase activity was then measured. (D) RHS5(WT)-IL4P(ΔOct1) (RHS5-IL4P construct in which Oct1 binding sites were deleted within the Il4 promoter) or RHS5(mtOct1)-IL4P(ΔOct1) (RHS5-IL4 reporter construct bearing both mutation at Oct-1 binding site within RHS5 and deletion of Oct-1 binding sites within the Il4 promoter) were transfected into EL4 cells with expression vectors (Oct1-CMV, GATA-3-CMV, or control CMV) by electroporation. Luciferase activity was measured as (C). Data are representative of three independent experiments with similar results (NT = no treatment). Bars are shown to indicate mean ± SD (n = 3). Statistical difference between groups was analyzed by Student’s t -test (*: P < 0.05).

Article Snippet: The T lymphocyte EL4 cell line and the Phoenix-Eco cell line were purchased from the Korean Cell Line Bank (Laboratory of Cell Biology, Cancer Research Center and Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea).

Techniques: Transfection, Expressing, Control, Construct, Electroporation, Luciferase, Activity Assay, Mutagenesis, Binding Assay