jagged1 fc fusion protein  (R&D Systems)

Journal: Cell Death Discovery

Article Title: Circulating mature dendritic cells homing to the thymus promote thymic epithelial cells involution via the Jagged1/Notch3 axis

doi: 10.1038/s41420-021-00619-5

Figure Lengend Snippet: A BMDCs were stained with the anti-Jagged1 antibody. The expression level of Jagged1 in BMDCs was examined using FCM after stimulation with different concentrations of LPS for 6 h. B The expression level of Jagged1 in BMDCs after stimulation with LPS (100 ng/ml) for different durations was detected using FCM. C Representative images of the expression of Notch3 (green fluorescence) and cytokeratin 8 (K8, a cTEC marker, red fluorescence) in the thymus from different aged mice (2–3 weeks old, 6–8 weeks old, and 10 months old, respectively) were captured by immunofluorescence microcopy. Paraffin sections of the thymus obtained from different aged mice were stained with primary rabbit anti-Notch3 and rat anti-cytokeratin 8 antibodies, followed by secondary Alexa Fluor 488-labeled goat anti-rabbit and Alexa Fluor 555-labeled goat anti-rat antibodies, respectively. Finally, the sections were stained with DAPI. Scale bars represent 200 µm. D mTEC1 cells were stained with PE-conjugated anti-Notch3 antibody. The expression of Notch3 in mTEC1 cells was detected by FCM. All data are from three independent experiments. Representative figures of these three experiments are displayed.

Article Snippet: The recombinant mouse Jagged1-human Fc fusion protein (rmJagged1-hFc) was procured from Chimerigen Laboratories (Cat#CHI-MF-111JAG1, MA, USA).

Techniques: Staining, Expressing, Fluorescence, Marker, Immunofluorescence, Labeling

Journal: Cell Death Discovery

Article Title: Circulating mature dendritic cells homing to the thymus promote thymic epithelial cells involution via the Jagged1/Notch3 axis

doi: 10.1038/s41420-021-00619-5

Figure Lengend Snippet: A rmJagged1-hFc was precoated on a 24-well plate at 4 °C for 12 h. BSA precoated at the same concentration was used as the control. mTEC1 cells were seeded at a density of 6 × 10 4 cells per well. Cell apoptosis was examined through staining with Annexin V/PI. B L and J cells were pre-stained with CFSE. mTEC1 cells were then cocultured with L or J cells at a 1:3 ratio for 48 h. The apoptosis of mTEC1 cells was examined through staining with Annexin V-APC/7-AAD. C The different concentrations of rmJagged1-hFc were precoated on 96-well plates at 4 °C for 12 h. The same concentration of BSA was set as the control group. mTEC1 cells were seeded at a density of 5 × 10 3 cells per well. Cell proliferation was examined using an EdU incorporation assay after 48 h. The red fluorescent dots indicate proliferating cells. Scale bars represent 200 µm. D The percentages of red fluorescent dots in all cells were calculated according to panel C . E Experiments were performed as described in A . mTEC1 cells were treated with or without DAPT (10 μM). Cell apoptosis was analyzed through Annexin V/PI staining. F Experiments were performed as described in C . Proliferation was detected by performing an EdU incorporation assay. mTEC1 cells were incubated with or without DAPT (10 μM). Red fluorescent dots indicate proliferating cells. Scale bars represent 200 µm. G The percentages of red fluorescent dots in all cells were calculated according to panel F . H Experiments were performed as described in B . mTEC1 cells were treated with or without DAPT (10 μM). I Experiments were performed as shown in Fig. . mTEC1 cells were cultured with or without DAPT (10 μM). J Experiments were performed as shown in Fig. . mTEC1 cells were treated with or without DAPT (10 μM). Scale bars represent 200 µm. K The percentages of red fluorescent dots in all cells were calculated according to panel J . All data are from three independent experiments. Representative figures of these three experiments are shown. Data are expressed as the mean ± SD. ** P < 0.01, *** P < 0.001 compared with the control group.

Article Snippet: The recombinant mouse Jagged1-human Fc fusion protein (rmJagged1-hFc) was procured from Chimerigen Laboratories (Cat#CHI-MF-111JAG1, MA, USA).

Techniques: Concentration Assay, Staining, Incubation, Cell Culture

Journal: Cell Death Discovery

Article Title: Circulating mature dendritic cells homing to the thymus promote thymic epithelial cells involution via the Jagged1/Notch3 axis

doi: 10.1038/s41420-021-00619-5

Figure Lengend Snippet: For A to F : Thymic atrophy after intrathymic injection of rmJagged1-hFc ( n = 3). The same volume of PBS was used as a control ( n = 3). A Thymus morphology and size. B Total numbers of thymocytes were calculated. C Thymocytes were stained with PE-conjugated anti-CD4 and APC-labeled anti-CD8 antibodies. The thymocyte subpopulations were analyzed by FCM. D The percentages and absolute numbers of the indicated thymocyte populations were calculated. E The cells isolated from the thymus of recipient mice were stained with Alexa Fluor 488-conjugated anti-CD45 and APC-labeled anti-EpCAM antibodies, respectively. CD45 - EpCAM + TECs were evaluated by FCM. F The percentages and absolute numbers of CD45 - EpCAM + cells were calculated. All data are from three independent experiments. Representative figures of these three experiments are shown. Data are represented as the mean ± SD. * P < 0.05, *** P < 0.001 compared with the control group.

Article Snippet: The recombinant mouse Jagged1-human Fc fusion protein (rmJagged1-hFc) was procured from Chimerigen Laboratories (Cat#CHI-MF-111JAG1, MA, USA).

Techniques: Injection, Staining, Labeling, Isolation

Journal: Cell Death Discovery

Article Title: Circulating mature dendritic cells homing to the thymus promote thymic epithelial cells involution via the Jagged1/Notch3 axis

doi: 10.1038/s41420-021-00619-5

Figure Lengend Snippet: Circulating mDCs can return to the thymus where they interact with TECs through direct cell–cell contact. MDCs can activate Notch signaling in TECs through the Jagged1/Notch3 axis. Long-term Notch signaling activation of TECs results in apoptosis and growth inhibition, further leading to the degeneration of the thymus.

Article Snippet: The recombinant mouse Jagged1-human Fc fusion protein (rmJagged1-hFc) was procured from Chimerigen Laboratories (Cat#CHI-MF-111JAG1, MA, USA).

Techniques: Activation Assay, Inhibition

Journal: Data in Brief

Article Title: RNA sequencing data of Notch ligand treated human dental pulp cells

doi: 10.1016/j.dib.2018.01.058

Figure Lengend Snippet: Information of samples for differential gene expression of RNA sequencing analysis of indirect immobilized Jagged1 treated human dental pulp cells.

Article Snippet: For Jagged1 treatment, recombinant human Jagged1/Fc fusion protein (10 nM; R&D systems, Minneapolis, MN, USA) was indirectly immobilized on tissue culture surfaces according to a previously published protocol .

Techniques: Gene Expression, RNA Sequencing, RNA Extraction, Recombinant

Journal: Data in Brief

Article Title: RNA sequencing data of Notch ligand treated human dental pulp cells

doi: 10.1016/j.dib.2018.01.058

Figure Lengend Snippet: Information of samples for differential gene expression of RNA sequencing analysis of indirect immobilized Jagged1 treated human dental pulp cells.

Article Snippet: For Jagged1 treatment, recombinant human Jagged1/Fc fusion protein (10 nM; R&D systems, Minneapolis, MN, USA) was indirectly immobilized on tissue culture surfaces according to a previously published protocol .

Techniques: Expressing, RNA Sequencing Assay, RNA Extraction, Recombinant

Journal: Data in Brief

Article Title: RNA sequencing data of Notch ligand treated human dental pulp cells

doi: 10.1016/j.dib.2018.01.058

Figure Lengend Snippet: Quality check of input total RNA using the Bioanalyzer. (A-C) hFc replicates; (D-F) Jagged1 replicates; (G-I) Jagged1+DAPT replicates.

Article Snippet: For Jagged1 treatment, recombinant human Jagged1/Fc fusion protein (10 nM; R&D systems, Minneapolis, MN, USA) was indirectly immobilized on tissue culture surfaces according to a previously published protocol .

Techniques:

Journal: Data in Brief

Article Title: RNA sequencing data of Notch ligand treated human dental pulp cells

doi: 10.1016/j.dib.2018.01.058

Figure Lengend Snippet: Library quality and size check using the Bioanalyzer. (A-C) hFc replicates; (D-F) Jagged1 replicates; (G-I) Jagged1+DAPT replicates.

Article Snippet: For Jagged1 treatment, recombinant human Jagged1/Fc fusion protein (10 nM; R&D systems, Minneapolis, MN, USA) was indirectly immobilized on tissue culture surfaces according to a previously published protocol .

Techniques:

Journal: Data in Brief

Article Title: RNA sequencing data of Notch ligand treated human dental pulp cells

doi: 10.1016/j.dib.2018.01.058

Figure Lengend Snippet: Average library size and concentration.

Article Snippet: For Jagged1 treatment, recombinant human Jagged1/Fc fusion protein (10 nM; R&D systems, Minneapolis, MN, USA) was indirectly immobilized on tissue culture surfaces according to a previously published protocol .

Techniques: Concentration Assay

Journal: Data in Brief

Article Title: RNA sequencing data of Notch ligand treated human dental pulp cells

doi: 10.1016/j.dib.2018.01.058

Figure Lengend Snippet: RNA-Seq alignment summary.

Article Snippet: For Jagged1 treatment, recombinant human Jagged1/Fc fusion protein (10 nM; R&D systems, Minneapolis, MN, USA) was indirectly immobilized on tissue culture surfaces according to a previously published protocol .

Techniques:

Journal: Data in Brief

Article Title: RNA sequencing data of Notch ligand treated human dental pulp cells

doi: 10.1016/j.dib.2018.01.058

Figure Lengend Snippet:

Article Snippet: For Jagged1 treatment, recombinant human Jagged1/Fc fusion protein (10 nM; R&D systems, Minneapolis, MN, USA) was indirectly immobilized on tissue culture surfaces according to a previously published protocol .

Techniques: High Throughput Screening Assay, RNA Sequencing Assay, Control, Isolation, Sequencing, Expressing