Journal: iScience

Article Title: The proprotein convertase furin regulates the development of thymic epithelial cells to ensure central immune tolerance

doi: 10.1016/j.isci.2022.105233

Figure Lengend Snippet: Furin promoted the development and proliferation of TECs via proteolytic cleavage of proIGF1R and proIR (A) A row-normalized heatmap showing the expression of gene pathways that gradually decreased in mTECs and cTECs of 2-week-old WT, Furin fl/+ Foxn1 Cre , and Furin fl/fl Foxn1 Cre mice. (B) UMAP plots (left) and violin plots (right) showing the expression of Igf1r and Insr on mTECs and cTECs of 2-week-old WT and Furin fl/fl Foxn1 Cre mice. (C) Immunoblotting of cell lysates of cultured WT and Furin KO TECs for detecting the cleavage of proIGF1R and proIR. (D) Immunoblotting of cell lysates of cultured WT and Furin KO TECs for detecting the phosphorylation of IGF1R/IR, ERK1/2, and Akt (Thr308 and Ser473). (E) mTECs of WT and Furin fl/fl Foxn1 Cre mice were sorted by flow cytometry, and cell lysates were analyzed by Simple Western to detect the cleavage of proIGF1R and proIR. (F) Cultured WT and Furin KO TECs were starved overnight following stimulation with exogenous IGF1 (100 ng/mL) and insulin (200 ng/mL) for 20 min, and cell lysates were analyzed by immunoblot for detecting the phosphorylation of IGF1R/IR, ERK1/2, and Akt (Thr308 and Ser473). In panels C-F, the results are representative of three independent experiments. (G) The numbers of mTECs and cTECs in ERK inhibitor (SCH772984) treated mice (n = 4) and control mice (n = 5). (H) The numbers of mTECs and cTECs in Akt inhibitor (MK-2206) treated fetal thymic lobes (n = 6) and control fetal thymic lobes (n = 7). Data are represented as mean ± SD ∗p < 0.05; ∗∗p < 0.01; NS, not statistically different.

Article Snippet: The primary antibodies used for Simple Western are as follows: β-Actin (Sigma-Aldrich, A5441), IGF1R antibody (Novus Biologicals, NBP1-77679), and Insulin receptor β (CST, 3025).

Techniques: Expressing, Western Blot, Cell Culture, Phospho-proteomics, Flow Cytometry, Simple Western, Control

Journal: iScience

Article Title: The proprotein convertase furin regulates the development of thymic epithelial cells to ensure central immune tolerance

doi: 10.1016/j.isci.2022.105233

Figure Lengend Snippet:

Article Snippet: The primary antibodies used for Simple Western are as follows: β-Actin (Sigma-Aldrich, A5441), IGF1R antibody (Novus Biologicals, NBP1-77679), and Insulin receptor β (CST, 3025).

Techniques: Plasmid Preparation, Recombinant, Reverse Transcription, Bicinchoninic Acid Protein Assay, Software