Journal: iScience

Article Title: Uhrf1 governs the proliferation and differentiation of muscle satellite cells

doi: 10.1016/j.isci.2022.103928

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-Myh3 , DSHB , cat# F1.652; RRID: AB_528358.

Techniques: Recombinant, Western Blot, Imaging, Multiplex Assay, Library Quantification, Methylation, Electrophoresis, Software

Journal: Oncogenesis

Article Title: Post-transcriptional screen of cancer amplified genes identifies ERBB2 / Her2 signaling as AU-rich mRNA stability-promoting pathway

doi: 10.1038/s41389-021-00351-w

Figure Lengend Snippet: A The ERBB2-amplification associated expressed genes (≥1.7 fold, Q value <0.00005) were intersected with the AU-rich element database (ARED-Plus). B Functional enrichment of the ERBB2-associated ARE-coding genes. The most significant results with false discovery rate (FDR) P < 0.05 are shown. C Left panel: the mean of mRNA abundance for 160 ARE-coding genes (Fig. data) was compared between TCGA ERBB2-negative and ERBB2-positive patients’ tissue samples. Data are Mean ± SEM. *** p < 0.001 using Student’s t-test with Welch’s correction. Right panel: the interaction between the ARE-mRNA cluster or all mRNAs with ERBB2-positive gene expression data is represented by an odd ratio using Fisher’s exact test (Oncomine software). D Total RNA from the ERBB2-positive SKBR3 cell line and MCF10A normal-like ERBB2-negative cell lines were extracted and subjected to RT-QPCR using specific primers to the indicated genes. Inset shows an immunoblot of two for the ERBB2 protein among different types of breast cancer cell lines. E The SKBR3 cells were transfected with 200 nM control siRNA or ERBB2 siRNA for 24 h, followed by RNA extraction and RT-QPCR as indicated. Inset is a representative immunoblot of three, for the ERBB2 protein. The data in D and E are normalized to the housekeeping gene, RPL0. Data are Mean ± SEM (triplicate) from one representative experiment of at least two. ANOVA test demonstrates overall statistical significance. The student’s t-test was used to compare expression data for each gene.

Article Snippet: The human breast ERBB2-expressing adenocarcinoma cell lines SKBR3 and BT-474, ERBB2-negative MCF-7 cell line, breast normal-like cell line MCF10A, and the Human embryonic kidney cells (HEK293) cell lines were obtained from American Type Culture Collection (ATCC; Rockville, MD).

Techniques: Amplification, Functional Assay, Gene Expression, Software, Quantitative RT-PCR, Western Blot, Transfection, Control, RNA Extraction, Expressing

Journal: Oncogenesis

Article Title: Post-transcriptional screen of cancer amplified genes identifies ERBB2 / Her2 signaling as AU-rich mRNA stability-promoting pathway

doi: 10.1038/s41389-021-00351-w

Figure Lengend Snippet: A SKBR3 cells were treated with increasing doses of lapatinib for 24 h, and lysed for immunoblotting with anti-phospho ERBB2 protein and β-actin as a loading control. WB is one representative from three blots. B RT-QPCR expression analysis for the ERBB2-regulated ARE-mRNAs in SKBR3 cells (left panel) or MCF-10A cells ( right panel ) treated with DMSO or Lapatinib (300 nM) for overnight. Data are normalized to the housekeeping gene, RPL0. The normalized values were used to calculate % remaining relative to DMSO control. Statistical analysis was performed using two-tailed Student’s t -test for each gene reduced in expression in comparison to DMSO data. Data are Mean ± SEM (triplicate of one experiment of three). ** P < 0.001, *** P < 0.0001. C RT-QPCR for Nanoluciferase mRNA abundance or Nanoluciferase protein activity ( D ) in SKBR3 cells transfected with nLuc-ARE or control nLuc-non-ARE expression plasmid for overnight, and then treated with DMSO (control) or lapatinib for 6 h or 24 h, respectively. Data are presented as the mean ± SEM ( n = 2 experiments). ANOVA is used to compare overall significance while post hoc sidak’s test compares the DMSO and lapatinib data. *** p < 0.0001. ** p < 0.005, Student’s test. E The mRNA half-life determination in response to lapatinib. The SKBR3 cells were treated with DMSO or lapatinib (300 nM) overnight, then actinomycin D (AcD) (5 μg/ml) was added for the indicated time points. Total RNA was extracted, and cDNA samples were subjected to RT-QPCR using specific primers to the indicated genes. One-phase exponential decay curves for relative mRNA half-life measurements in the two treated groups are shown. Data are normalized to RPL0 transcript abundance, and the normalized values were used to calculate the mRNA half-life as a percentage to time 0. Data are presented as the mean ± SEM ( n = 2 experiments). F , G Immunoblot analysis for CDC6 and NEK2 protein abundance in SKBR3 cells treated as in ( A ). Representative blot each is from at least two experiments is shown.

Article Snippet: The human breast ERBB2-expressing adenocarcinoma cell lines SKBR3 and BT-474, ERBB2-negative MCF-7 cell line, breast normal-like cell line MCF10A, and the Human embryonic kidney cells (HEK293) cell lines were obtained from American Type Culture Collection (ATCC; Rockville, MD).

Techniques: Western Blot, Control, Quantitative RT-PCR, Expressing, Two Tailed Test, Comparison, Activity Assay, Transfection, Plasmid Preparation, Quantitative Proteomics

Journal: Oncogenesis

Article Title: Post-transcriptional screen of cancer amplified genes identifies ERBB2 / Her2 signaling as AU-rich mRNA stability-promoting pathway

doi: 10.1038/s41389-021-00351-w

Figure Lengend Snippet: A immunoblot analysis for ZFP36/TTP phosphorylated protein abundance in MCF10A and SKBR3 using specific primers to ZFP36/TTP and β-Actin protein as a loading control. B SKBR3 cells were transfected with an empty vector or ZFP36/TTP expression vector. Left panel: proteins were extracted, and immunoblotting was performed using a ZFP36/TTP antibody. Right panel: protein lysates were first treated with the phosphatase, CIP, or CIP phosStop C before immunoblotting with anti-ZFP36/TTP. The WB shows both phosphorylated and unphosphorylated bands. WB was performed with low exposure to avoid high background and overexposed signal of the transfected TTP (lower panel). D) SKBR3 cells were transfected with 200 nM ERBB2 siRNA or control siRNA for overnight, and ERBB2 and ZFP36/TTP protein abundance were assessed by immunoblotting. A representative blot from at least two experiments is shown. Immunoblot analysis of ERBB2 and ZFP36/TTP protein phosphorylation abundance in MCF10A ( E ) or MCF-7 ( F ) transfected with ERBB2 expression vector or empty vector. Representative blots are from at least three experiments.

Article Snippet: The human breast ERBB2-expressing adenocarcinoma cell lines SKBR3 and BT-474, ERBB2-negative MCF-7 cell line, breast normal-like cell line MCF10A, and the Human embryonic kidney cells (HEK293) cell lines were obtained from American Type Culture Collection (ATCC; Rockville, MD).

Techniques: Western Blot, Quantitative Proteomics, Control, Transfection, Plasmid Preparation, Expressing, Phospho-proteomics

Journal: Oncogenesis

Article Title: Post-transcriptional screen of cancer amplified genes identifies ERBB2 / Her2 signaling as AU-rich mRNA stability-promoting pathway

doi: 10.1038/s41389-021-00351-w

Figure Lengend Snippet: A Dose-dependent activity of lapatinib on the abundance of phosphorylated ZFP36/TTP in the ERBB2-positive SKBR3 cells. The relative amounts of the proteins from three immunoblots were quantified using Image J software ( left panel ). Fold changes (Mean ± SEM, n = 3) of phosphorylated ZFP36/TTP abundance normalized to β-Actin using Image J program (right panel). B Effect of ERBB2 inhibition on ZFP36/TTP protein stability. SKBR3 cells were treated with lapatinib (300 nM) for 16 h, followed by 10 µg/ml of cycloheximide (CHX) treatment for the indicated durations (left panel). Protein lysates were subjected to immunoblotting with anti-TTP. Right panel: quantification of ZFP36/TTP abundance, normalized to GAPDH, was determined using ImageJ software and the protein half-life was assessed using the Graph Prism. Data are mean +/− SEM of two experiments. C Effect of lapatinib on the abundance of ZFP36/TTP protein in the ERBB2-positive cells, BT-474. WB is a representative of two blots from independent experiments.

Article Snippet: The human breast ERBB2-expressing adenocarcinoma cell lines SKBR3 and BT-474, ERBB2-negative MCF-7 cell line, breast normal-like cell line MCF10A, and the Human embryonic kidney cells (HEK293) cell lines were obtained from American Type Culture Collection (ATCC; Rockville, MD).

Techniques: Activity Assay, Western Blot, Software, Inhibition

Journal: Oncogenesis

Article Title: Post-transcriptional screen of cancer amplified genes identifies ERBB2 / Her2 signaling as AU-rich mRNA stability-promoting pathway

doi: 10.1038/s41389-021-00351-w

Figure Lengend Snippet: A Immunoblot analysis of ERBB2, AKT, MEK1/2, and ERK1/2 signaling inhibition after treatment with increasing doses of lapatinib for 24 h. Status of ERBB2 (phospho-tyrosine 1221/1222 and total), AKT (phospho-serine 473 and total pan AKT), ERK1/2 (Phospho-threonine 202/phospho-tyrosine 204 and total), MEK1/2 (Ser217/221), and ZFP36/TTP are shown. A blot from at least two experiments is shown. B Immunoblot analysis for the inhibition of MK2 activation after treatment with lapatinib. SKBR3 cells were treated with increasing doses of lapatinib, as indicated for 24 h. Phospho-threonine MK2 (T222) and total MK2 protein abundance are shown. A representative blot from two experiments is shown. C Immunoblot analysis for ZFP36/TTP phosphorylation in MAPKAPK-2 silenced SKBR3 cells. SKBR3 cells transfected with 100 nM of si Ctrl or si MAPKAPK-2 for 24 h were tested for ZFP36/TTP protein abundance. Total MK2, phosphorylated ZFP36/TTP, and β-actin protein abundances are shown. A representative blot from three experiments is shown. Effect of the MK2 inhibitor PF-3644022 (1 µM, 4 h) on the abundance of phosphorylated ZFP36/TTP in ERBB2-expressing SKBR3 ( D ) or SKBR3 transfected with ZFP36 expression plasmid ( E ). Western blots are from two independent experiments. F Immunoblot analysis of total and phosphorylated ZFP36/TTP, and MK2 in MK2 -knockout MEF cells in the absence or presence of overnight transfected ERBB2.

Article Snippet: The human breast ERBB2-expressing adenocarcinoma cell lines SKBR3 and BT-474, ERBB2-negative MCF-7 cell line, breast normal-like cell line MCF10A, and the Human embryonic kidney cells (HEK293) cell lines were obtained from American Type Culture Collection (ATCC; Rockville, MD).

Techniques: Western Blot, Inhibition, Activation Assay, Quantitative Proteomics, Phospho-proteomics, Transfection, Expressing, Plasmid Preparation, Knock-Out

Journal: eLife

Article Title: Fine-tuning of β-catenin in mouse thymic epithelial cells is required for postnatal T-cell development

doi: 10.7554/eLife.69088

Figure Lengend Snippet: ( A ) Thoracic cavity of control mice and β-cat gain-of-function (GOF) mice at E15.5. Dotted lines show the outline of the thymic primordium. In many cases, blood clots were observed in the central core of the thymic primordium in β-cat GOF mice. Representative data from three independent experiments are shown. TH: thymus, H: heart. Bar: 1 mm. ( B ) Labeling of β5t-iCre-activated cell progenies with tdTomato fluorescence in the thymus of control mice and β-cat GOF mice at E15.5. Mouse sections were nuclear counterstained with TO-PRO3. Bottom panels are magnifications of white-boxed areas in the top panels. Representative data from three independent experiments are shown. Bars: indicated in figures. ( C ) Hematoxylin and eosin staining (top) and immunofluorescence staining for K5 and K8 (middle) and β-catenin and β5t (bottom) on sagittal sections of thymic primordium in control mice and β-cat GOF mice at E15.5. Representative results from three independent experiments are shown. Bar: 100 μm. ( D ) Intracellular staining of β-catenin in CD45 − EpCAM + TECs isolated from control mice and β-cat GOF mice at E15.5. Histograms show β-catenin expression in control TECs (blue line) and β-cat GOF TECs (red line). Shaded area and black line represent the fluorescence in the absence of anti-β-catenin antibody in control TECs and β-cat GOF TECs, respectively. Plots on the right show net median fluorescence intensity (MFI) values (means and standard error of the means [SEMs], n = 4–5). The numbers in parentheses indicate percentage of control value. *p < 0.05.

Article Snippet: For the isolation of TECs, CD45 − cells were enriched with magnetic-bead-conjugated anti-CD45 antibody (Miltenyi Biotec) before multicolor staining for flow cytometric cell sorting.

Techniques: Control, Labeling, Fluorescence, Staining, Immunofluorescence, Isolation, Expressing

Journal: eLife

Article Title: Fine-tuning of β-catenin in mouse thymic epithelial cells is required for postnatal T-cell development

doi: 10.7554/eLife.69088

Figure Lengend Snippet: ( A ) Immunofluorescence staining for CD45 and Foxn1 on sagittal sections of thymic primordium in control mice and β-cat GOF mice at E11.5–E15.5. The sections were nuclear counterstained with TO-PRO3. Anterior–posterior (A–P) and dorsal–ventral (D–V) orientations of the images are indicated. Representative data from three independent experiments are shown. Bar: 100 μm. ( B ) Quantitative RT-PCR analysis of mRNA expression levels (means and standard error of the means [SEMs], n = 3–4) of indicated genes relative to Gapdh levels in CD45 − EpCAM + thymic epithelial cells (TECs) isolated from the thymus of control mice and β-cat GOF mice at E15.5. ( C ) The numbers of CD45 + thymocytes were analyzed by flow cytometry. Plots show the numbers (means and SEMs, n = 3–4) of CD45 + thymocytes in the thymus of control mice and β-cat GOF mice at E15.5. ( D ) Flow cytometric analysis of double negative (DN) thymocytes from control mice and β-cat GOF mice at E15.5. Shown are profiles of CD44 and CD25 expression. The numbers in dot plots indicate the frequency of cells within indicated area. ( E ) Cell numbers (means and SEMs, n = 3–4) of indicated DN thymocyte subpopulations from control mice and β-cat GOF mice at E15.5 are plotted. *p < 0.05; **p < 0.01; ***p < 0.001; N.S., not significant.

Article Snippet: For the isolation of TECs, CD45 − cells were enriched with magnetic-bead-conjugated anti-CD45 antibody (Miltenyi Biotec) before multicolor staining for flow cytometric cell sorting.

Techniques: Immunofluorescence, Staining, Control, Quantitative RT-PCR, Expressing, Isolation, Flow Cytometry

Journal: eLife

Article Title: Fine-tuning of β-catenin in mouse thymic epithelial cells is required for postnatal T-cell development

doi: 10.7554/eLife.69088

Figure Lengend Snippet: ( A ) Immunofluorescence staining for CD45 and Foxn1 on sagittal sections of the thymic primordium in control mice and β-cat gain-of-function (GOF) mice at E11.5–E15.5. Shown are merged images with nuclear counterstaining (TO-PRO3) (left) and images obtained in each channel (middle, right) for the thymus from control mice and β-cat GOF mice at the indicated stages. Representative data from three independent experiments are shown. Bar: 100 μm. ( B ) Flow cytometric analysis of thymic epithelial cells (TECs) from indicated mice at E15.5, showing the purity of the isolated TECs for quantitative RT-PCR analysis. Shown are representative profiles of CD45, PI, and EpCAM expression in total cells (left) and in isolated CD45 − EpCAM + TECs (right) from control mice and β-cat loss-of-function (LOF) mice. The numbers indicate the frequency of cells within indicated areas.

Article Snippet: For the isolation of TECs, CD45 − cells were enriched with magnetic-bead-conjugated anti-CD45 antibody (Miltenyi Biotec) before multicolor staining for flow cytometric cell sorting.

Techniques: Immunofluorescence, Staining, Control, Isolation, Quantitative RT-PCR, Expressing

Journal: eLife

Article Title: Fine-tuning of β-catenin in mouse thymic epithelial cells is required for postnatal T-cell development

doi: 10.7554/eLife.69088

Figure Lengend Snippet: ( A ) Flow cytometric analysis of splenocytes from control mice and β-cat gain-of-function (GOF) mice at 11 wk. Shown are representative dot plot profiles of CD3 and TCRβ expression (left) and CD3 and TCRδ expression (right) in PI − viable cells. The numbers in dot plots indicate the frequency of cells within indicated area. ( B ) Cell numbers (means and standard error of the means [SEMs], n = 4–6) of indicated subpopulations in the spleen (left) and the iLN (right) from control and β-cat GOF mice at 8 wk are plotted. ( C ) Shown are representative dot plots of TCRδ and Vγ5 expression in CD45 + cells in the thymus from control mice and β-cat GOF mice at E15.5. The frequency of TCRδ + Vγ5 + cells is plotted (mean and SEMs; n = 4–6). ( D ) Flow cytometric analysis of dendritic epidermal T cells (DETCs) in the skin epidermis from control mice and β-cat GOF mice at 8 wk. Shown are representative dot plot profiles of CD3 and Vγ5 expression in epidermal cells. The numbers in dot plots indicate the frequency of cells within indicated area. The frequency of CD3 + Vγ5 + cells in epidermal cells is plotted (means and SEMs; n = 3). ( E ) Histograms for Vγ4 and Vγ1 expression in TCRβ - δ + cells in the iLN from control mice and β-cat GOF mice at 8 wk are shown. The frequency of Vγ4 + and Vγ1 + cells is plotted (means and SEMs; n = 3). *p < 0.05; **p < 0.01; ***p < 0.001; N.S., not significant.

Article Snippet: For the isolation of TECs, CD45 − cells were enriched with magnetic-bead-conjugated anti-CD45 antibody (Miltenyi Biotec) before multicolor staining for flow cytometric cell sorting.

Techniques: Control, Expressing

Journal: eLife

Article Title: Fine-tuning of β-catenin in mouse thymic epithelial cells is required for postnatal T-cell development

doi: 10.7554/eLife.69088

Figure Lengend Snippet: ( A ) Thoracic cavity of control mice and β-cat LOF mice at E15.5. Dotted lines show the outline of the thymic primordium. Representative data from three independent experiments are shown. TH: thymus, H: heart. Bar: 1 mm. ( B ) Immunofluorescence staining for β-catenin and Foxn1 (left) or K5 and K8 (right) on sagittal sections of the thymus from control mice and β-cat LOF mice at E15.5. Shown are merged images with nuclear counterstaining (TO-PRO3) (top) and images obtained in each channel (middle, bottom). Representative data from three independent experiments are shown. Bar: 100 μm. ( C ) Intracellular staining of β-catenin in CD45 − EpCAM + thymic epithelial cells (TECs) from control mice and β-cat LOF mice at E15.5. Histograms show β-catenin expression in control TECs (blue line) and β-cat LOF TECs (red line). Shaded area and black line represent the fluorescence in the absence of anti-β-catenin antibody in control TECs and β-cat LOF TECs, respectively. Plots show net median fluorescence intensity (MFI) values for β-catenin (means and standard error of the means [SEMs], n = 4). The numbers in parentheses indicate percentage of control value. ( D ) Flow cytometric analysis of enzyme-digested thymic cells from indicated mice at E15.5. Shown are profiles of EpCAM and CD45 expression in PI − viable cells (left) and UEA1 reactivity and Ly51 expression in CD45 − EpCAM + cells (right). The numbers in dot plots indicate the frequency of cells within indicated area. ( E ) Plots show the number of total thymic cells (left) and the frequency and the number of total TECs (middle) and cortical thymic epithelial cell (cTECs; right) from control mice and β-cat LOF mice at E15.5 (means and SEMs, n = 4). **p < 0.01; N.S., not significant.

Article Snippet: For the isolation of TECs, CD45 − cells were enriched with magnetic-bead-conjugated anti-CD45 antibody (Miltenyi Biotec) before multicolor staining for flow cytometric cell sorting.

Techniques: Control, Immunofluorescence, Staining, Expressing, Fluorescence

Journal: eLife

Article Title: Fine-tuning of β-catenin in mouse thymic epithelial cells is required for postnatal T-cell development

doi: 10.7554/eLife.69088

Figure Lengend Snippet: ( A ) Intracellular staining of β-catenin in UEA1 − Ly51 + cortical thymic epithelial cells (cTECs; left) and UEA1 + Ly51 − medullary thymic epithelial cells (mTECs; right) from control mice and β-cat LOF mice at 2 wk. Histograms show β-catenin expression in cTECs and mTECs from control mice (blue line) and β-cat LOF mice (red line). Shaded area and black line represent the fluorescence in the absence of anti-β-catenin antibody in control TECs and β-cat LOF TECs, respectively. Plots show net MFI values for β-catenin in cTECs and mTECs (means and standard error of the means [SEMs], n = 3). The numbers in parentheses indicate percentage of control value. ( B ) Quantitative RT-PCR analysis of mRNA expression levels (means and SEMs, n = 5) of indicated genes relative to Gapdh levels in UEA1 − Ly51 + cTECs (top) and UEA1 + Ly51 − mTECs (bottom) in the thymus of control mice and β-cat LOF mice at 2 wk. ( C ) Flow cytometric analysis of enzyme-digested thymic cells from control mice and β-cat LOF mice at 2 wk. Shown are representative profiles of EpCAM and CD45 expression in PI − viable cells (left) and UEA1 reactivity and Ly51 expression in CD45 − EpCAM + viable cells (right). The numbers in dot plots indicate the frequency of cells within indicated area. ( D ) Plots show the number (means and SEMs, n = 6) of cTECs and mTECs in the thymus from control mice and β-cat LOF mice at 2 wk. ( E ) Immunofluorescence analysis of β5t (green), CCL21 (red), and Aire (cyan) on transverse sections of thymus from control mice and β-cat LOF mice at 2 wk. Representative data from three independent experiments are shown. Bar: 100 μm. Ctrl: Control, LOF: β-cat LOF. *p < 0.05; **p < 0.01; ***p < 0.001; N.S., not significant.

Article Snippet: For the isolation of TECs, CD45 − cells were enriched with magnetic-bead-conjugated anti-CD45 antibody (Miltenyi Biotec) before multicolor staining for flow cytometric cell sorting.

Techniques: Staining, Control, Expressing, Fluorescence, Quantitative RT-PCR, Immunofluorescence

Journal: eLife

Article Title: Fine-tuning of β-catenin in mouse thymic epithelial cells is required for postnatal T-cell development

doi: 10.7554/eLife.69088

Figure Lengend Snippet: Flow cytometric analysis of cTECs and mTECs from indicated mice at 2 wk. Shown are representative profiles of CD45, PI, and EpCAM expression of total cells after magnetic depletion of CD45 + cells, UEA1 reactivity and Ly51 expression in CD45 − EpCAM + cells, and isolated cTECs and isolated mTECs from control mice and β-cat loss-of-function (LOF) mice. The numbers indicate the frequency of cells within indicated areas.

Article Snippet: For the isolation of TECs, CD45 − cells were enriched with magnetic-bead-conjugated anti-CD45 antibody (Miltenyi Biotec) before multicolor staining for flow cytometric cell sorting.

Techniques: Expressing, Isolation, Control

Journal: eLife

Article Title: Fine-tuning of β-catenin in mouse thymic epithelial cells is required for postnatal T-cell development

doi: 10.7554/eLife.69088

Figure Lengend Snippet: ( A ) Bars show body weight (left) and thymus weight (right) at 6 mo in control female mice and β-cat LOF female mice (means and standard error of the means [SEMs], n = 4). ( B ) Appearance of thymus from control mice and β-cat LOF mice at 6 mo. Representative data from four independent experiments are shown. Bar: 1 mm. ( C ) Plots show the number (means and SEMs, n = 4) of total thymocytes in the thymus from control mice and β-cat LOF mice at 6 mo. ( D ) Flow cytometric analysis of enzyme-digested thymic cells from control mice and β-cat LOF mice at 6 mo. Shown are representative profiles of EpCAM and CD45 expression in PI − viable cells (left) and UEA1 reactivity and Ly51 expression in CD45 − EpCAM + viable cells (right). The numbers in dot plots indicate the frequency of cells within indicated area. ( E ) Plots show the number (means and SEMs, n = 4) of cortical thymic epithelial cells (cTECs) and medullary thymic epithelial cells (mTECs) in the thymus from control mice and β-cat LOF mice at 6 mo. ( F ) Immunofluorescence analysis of K5 (green) and K8 (magenta) on transverse sections of thymus from control mice and β-cat LOF mice at 6 mo. Representative data from three independent experiments are shown. Bar: 100 μm. Ctrl: Control, LOF: β-cat LOF. *p < 0.05; N.S., not significant.

Article Snippet: For the isolation of TECs, CD45 − cells were enriched with magnetic-bead-conjugated anti-CD45 antibody (Miltenyi Biotec) before multicolor staining for flow cytometric cell sorting.

Techniques: Control, Expressing, Immunofluorescence

Journal: eLife

Article Title: Fine-tuning of β-catenin in mouse thymic epithelial cells is required for postnatal T-cell development

doi: 10.7554/eLife.69088

Figure Lengend Snippet:

Article Snippet: For the isolation of TECs, CD45 − cells were enriched with magnetic-bead-conjugated anti-CD45 antibody (Miltenyi Biotec) before multicolor staining for flow cytometric cell sorting.

Techniques: Transduction, Plasmid Preparation, Recombinant, Sequencing, DNA Library Preparation, Software