Journal: Frontiers in Medicine

Article Title: IL-7-Treated Periodontal Ligament Cells Regulate Local Immune Homeostasis by Modulating Treg/Th17 Cell Polarization

doi: 10.3389/fmed.2022.754341

Figure Lengend Snippet: Effects of conditioned medium of human periodontal ligament cells (hPDLCs) treated with rhIL-7 on the differentiation of Treg/Th17 cells. The supernatants of hPDLCs were collected after being treated with different concentrations of rhIL-7 for 24 h and used to culture CD4 + T cells for 3 days. Flow cytometry (FCM) was used to detect the polarization of CD4 + T cells into Treg cells (A,B) and Th17 cells (D,E) . (C) The concentration of Foxp3 in CD4 + T cells was detected by ELISA. (F) The ratio of Treg/Th17. Representative dot plots show the gating strategy of FCM analysis. n = 3, * p < 0.05. ** p < 0.01. ## DAPT group compared with 200 ng/mL rhIL-7 group, p < 0.01.

Article Snippet: About 1 × 10 5 hPDLCs were seeded in 6-well plates, and a fresh medium was introduced and supplemented with different concentrations of rhIL-7 (PeproTech, USA) after 24 h; cultivating was continued for another 24 h. Cells in the inhibitor (DAPT) group are pre-treated for 2 h at 37°C with 100 μM DAPT (MedChemExpress, USA) and then replaced with fresh medium supplemented with 200 ng/ml rhIL-7.

Techniques: Flow Cytometry, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Medicine

Article Title: IL-7-Treated Periodontal Ligament Cells Regulate Local Immune Homeostasis by Modulating Treg/Th17 Cell Polarization

doi: 10.3389/fmed.2022.754341

Figure Lengend Snippet: Primary human periodontal ligament cells. (A) Under the microscope, it can be seen that many fibroblast cells are crawling out around the dark brown primary tissue block. The cells are adherent to the wall and grow with a clear outline. Most of the cells are flat and spindle-shaped or star-shaped structures arranged in radial or whirlpool shape. Effect of rhIL-7 on the proliferation of hPDLCs. Scale bar = 1,000 μm. (B) After hPDLCs were treated with different concentrations of rhIL-7 for 24 h, CCK-8 was used to detect the cell survival rate of the hPDLCs. A summary of the results is shown in the bar graphs. Effect of rhIL-7 on the apoptosis and necrosis of hPDLCs. (C,D) We used Annexin V-FITC/PI staining to detect the proportion of apoptosis and necrosis in the cells of each group by FCM. Representative dot plots show the gating strategy of FCM analysis. The number in the scatter plot represents the percentage of cells in the corresponding state. n = 3, * p < 0.05. ** p < 0.01. CCK-8, cell counting kit-8; FCM, flow cytometry; hPDLCs, human periodontal ligament cells.

Article Snippet: About 1 × 10 5 hPDLCs were seeded in 6-well plates, and a fresh medium was introduced and supplemented with different concentrations of rhIL-7 (PeproTech, USA) after 24 h; cultivating was continued for another 24 h. Cells in the inhibitor (DAPT) group are pre-treated for 2 h at 37°C with 100 μM DAPT (MedChemExpress, USA) and then replaced with fresh medium supplemented with 200 ng/ml rhIL-7.

Techniques: Microscopy, Blocking Assay, CCK-8 Assay, Staining, Cell Counting, Flow Cytometry

Journal: Frontiers in Medicine

Article Title: IL-7-Treated Periodontal Ligament Cells Regulate Local Immune Homeostasis by Modulating Treg/Th17 Cell Polarization

doi: 10.3389/fmed.2022.754341

Figure Lengend Snippet: Effect of rhIL-7 on inflammatory factors related to the immune response in hPDLCs. (A–C) After hPDLCs were treated with different concentrations of rhIL-7 for 24 h, the concentration and protein level of TGF-β and IL-6 in the supernatants of each group were detected by ELISA and WB. n = 3, * p < 0.05. ** p < 0.01. # DAPT group compared with 200 ng/mL rhIL-7 group, p < 0.05. hPDLCs, human periodontal ligament cells; TGF, transforming growth factor; WB, Western blot.

Article Snippet: About 1 × 10 5 hPDLCs were seeded in 6-well plates, and a fresh medium was introduced and supplemented with different concentrations of rhIL-7 (PeproTech, USA) after 24 h; cultivating was continued for another 24 h. Cells in the inhibitor (DAPT) group are pre-treated for 2 h at 37°C with 100 μM DAPT (MedChemExpress, USA) and then replaced with fresh medium supplemented with 200 ng/ml rhIL-7.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Frontiers in Medicine

Article Title: IL-7-Treated Periodontal Ligament Cells Regulate Local Immune Homeostasis by Modulating Treg/Th17 Cell Polarization

doi: 10.3389/fmed.2022.754341

Figure Lengend Snippet: Effects of rhIL-7 on the expression PD-L1 in hPDLCs. (A–D) After hPDLCs were treated with different concentrations of rhIL-7 for 24 h, the MFI and the protein level of PD-L1 in each group were determined by FCM and WB. Representative histograms show the gating strategy of FCM analysis. n = 3, * p < 0.05. ** p < 0.01. ## DAPT group compared with 200 ng/mL rhIL-7 group, p < 0.01. FCM, flow cytometry; hPDLCs, human periodontal ligament cells; MFI, mean fluorescence intensity; WB, Western blot.

Article Snippet: About 1 × 10 5 hPDLCs were seeded in 6-well plates, and a fresh medium was introduced and supplemented with different concentrations of rhIL-7 (PeproTech, USA) after 24 h; cultivating was continued for another 24 h. Cells in the inhibitor (DAPT) group are pre-treated for 2 h at 37°C with 100 μM DAPT (MedChemExpress, USA) and then replaced with fresh medium supplemented with 200 ng/ml rhIL-7.

Techniques: Expressing, Flow Cytometry, Fluorescence, Western Blot

Journal: Frontiers in Medicine

Article Title: IL-7-Treated Periodontal Ligament Cells Regulate Local Immune Homeostasis by Modulating Treg/Th17 Cell Polarization

doi: 10.3389/fmed.2022.754341

Figure Lengend Snippet: Effect of rhIL-7 on signal molecules related to the immune response in hPDLCs. (A–C) The mRNA and protein levels of HIF-1α and TCF7 in hPDLCs were detected by RT-qPCR and WB after treatment with different concentrations of IL-7 for 24 h. A summary of the results is shown in the bar graphs. n = 3. ** p < 0.01. # DAPT group compared with 200 ng/mL rhIL-7 group, p < 0.05. ## DAPT group compared with 200 ng/mL rhIL-7 group, p < 0.01. HIF, hypoxia-inducible factor; hPDLCs, human periodontal ligament cells; RT-qPCR, reverse transcription quantitative PCR; WB, Western blot.

Article Snippet: About 1 × 10 5 hPDLCs were seeded in 6-well plates, and a fresh medium was introduced and supplemented with different concentrations of rhIL-7 (PeproTech, USA) after 24 h; cultivating was continued for another 24 h. Cells in the inhibitor (DAPT) group are pre-treated for 2 h at 37°C with 100 μM DAPT (MedChemExpress, USA) and then replaced with fresh medium supplemented with 200 ng/ml rhIL-7.

Techniques: Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Western Blot

Journal: Frontiers in Medicine

Article Title: IL-7-Treated Periodontal Ligament Cells Regulate Local Immune Homeostasis by Modulating Treg/Th17 Cell Polarization

doi: 10.3389/fmed.2022.754341

Figure Lengend Snippet: Effects of treatment with different concentrations of rhIL-7 in hPDLCs on Notch1 signaling pathway-related molecules. (A–C) After hPDLCs were treated with different concentrations of IL-7 for 24 h, the mRNA level and protein level of Notch1, Jagged1, and Hes1 were detected by RT-qPCR and WB. n = 3, * p < 0.05. ** p < 0.01. ## DAPT group compared with 200 ng/mL rhIL-7 group, p < 0.01. hPDLCs, human periodontal ligament cells; RT-qPCR, reverse transcription quantitative PCR; WB, Western blot.

Article Snippet: About 1 × 10 5 hPDLCs were seeded in 6-well plates, and a fresh medium was introduced and supplemented with different concentrations of rhIL-7 (PeproTech, USA) after 24 h; cultivating was continued for another 24 h. Cells in the inhibitor (DAPT) group are pre-treated for 2 h at 37°C with 100 μM DAPT (MedChemExpress, USA) and then replaced with fresh medium supplemented with 200 ng/ml rhIL-7.

Techniques: Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Western Blot

Journal: Developmental Cell

Article Title: Defining the identity and the niches of epithelial stem cells with highly pleiotropic multilineage potency in the human thymus

doi: 10.1016/j.devcel.2023.08.017

Figure Lengend Snippet:

Article Snippet: HumanKine® recombinant human IL-7 protein , Proteintech , cat# HZ-1281.

Techniques: Recombinant, Modification, Selection, RNA Extraction, Polymer, Cell Culture, Derivative Assay, Software