Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Impact of Fibronectin Knockout on Proliferation and Differentiation of Human Infrapatellar Fat Pad-Derived Stem Cells

doi: 10.3389/fbioe.2019.00321

Figure Lengend Snippet: Cell proliferation capacity in human IPFSCs after FN1-KO. FN1-KO cells were compared with copGFP in cell increase (A) , percentage of cells in the S and G 2 phases (B) , and surface markers [SSEA4 (C) , CD73 (D) , CD90 (E) , CD105 (F) , and CD146 (G) ] by flow cytometry; stemness genes ( NANOG, SOX2, KLF4, BMI1, MYC, NOV, POU5F1 , and NES ) (H) , senescent genes ( CDKN1A, CDKN2A , and TP53 ) (I) , and the mesenchymal condensation gene ( CDH2 ) (J) by qPCR. GAPDH was used as an endogenous control. Data are shown as bar charts. * indicates a significant difference compared to the corresponding copGFP group ( P < 0.05).

Article Snippet: The following primary antibodies were used: CD73-APC (cat no. 17-0739-42; eBioScience, Fisher Scientific, Waltham, MA), CD90-APC-Vio770 (cat no. 130-114-863; Miltenyi Biotec, San Diego, CA), CD105-PerCP-Vio700 (cat no. 130-112-170; Miltenyi Biotec), CD146-PE (cat no. 12-1469-42; eBioScience), and the stage-specific embryonic antigen 4-PE (SSEA4-PE; cat no. 330406; BioLegend, Dedham, MA).

Techniques: Flow Cytometry, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Impact of Fibronectin Knockout on Proliferation and Differentiation of Human Infrapatellar Fat Pad-Derived Stem Cells

doi: 10.3389/fbioe.2019.00321

Figure Lengend Snippet: Proliferation capacity of human IPFSCs after expansion on the dECMs deposited by FN1-KO cells. Passage 15 human IPFSCs were compared after expansion on dECMs deposited by Cas9-sgFN1a/b transduced cells (sgFN1a ECM and sgFN1b ECM, respectively) with those deposited by copGFP (copGFP ECM) and those grown on TCP (TCP) as controls in cell increase (A) , percentage of cells in the S and G 2 phases (B) , and surface markers [SSEA4 (C) , CD73 (D) , CD90 (E) , and CD105 (F) ] by flow cytometry; stemness genes ( NANOG, SOX2, KLF4, BMI1, MYC, NOV, POU5F1 , and NES ) (G) , senescence genes ( CDKN1A, CDKN2A , and TP53 ) (H) , and the mesenchymal condensation gene ( CDH2 ) (I) by qPCR. GAPDH was used as an endogenous control. Data are shown as bar charts. *indicates a significant difference compared to the corresponding copGFP group ( P < 0.05).

Article Snippet: The following primary antibodies were used: CD73-APC (cat no. 17-0739-42; eBioScience, Fisher Scientific, Waltham, MA), CD90-APC-Vio770 (cat no. 130-114-863; Miltenyi Biotec, San Diego, CA), CD105-PerCP-Vio700 (cat no. 130-112-170; Miltenyi Biotec), CD146-PE (cat no. 12-1469-42; eBioScience), and the stage-specific embryonic antigen 4-PE (SSEA4-PE; cat no. 330406; BioLegend, Dedham, MA).

Techniques: Flow Cytometry, Control

Journal: Nature

Article Title: AIM2 in regulatory T cells restrains autoimmune diseases.

doi: 10.1038/s41586-021-03231-w

Figure Lengend Snippet: Fig. 4 | AIM2 promotes Treg cells in vitro and restrains AKT–mTOR via the RACK1–PP2A complex. a, b, RT–PCR (a) and flow cytometry (b) of FOXP3 in wild-type and Aim2−/− CD4+ T cells activated with the indicated amounts (a) or 2 ng ml−1 (b) of TGFβ for 4 days; n = 5 experiments in a and n = 4 experiments in b. c, Flow cytometry of IFNγ+ or IL-17A+ CD4+ T cells of indicated genotypes, 4 days after differentiation under indicated polarizing conditions; n = 4 experiments. cTH17, classic TH17; pTH17, pathogenic TH17. d–g, Wild-type and Aim2−/− CD4+ T cells were stimulated as in b. d, Enrichment scores of indicated gene sets, based on RNA-seq datasets. e, f, ECAR (e) and OCR (f) levels during glycolysis, and OCR levels during fatty acid oxidation (g), by Seahorse analysis; n = 10 experiments for e; n = 5 experiments for f; and n = 3 experiments for g. h, Immunoblotting of indicated proteins in wild-type and Aim2−/− CD4+ T cells stimulated as in b with indicated treatment for 24 h. Representative of three

Article Snippet: Ectopic expression of PP2A and RACK1 in Treg cells To generate the retrovirus expressing PP2A and RACK1, we first cloned PP2A (OriGene Technologies, MR204384L4) and RACK1 (OriGene Technologies, MR204575L3) into retroviral vectors MSCV-IRES-Thy1.1 (MIT, Addgene 17442) and MSCV-IRES-GFP (MIG, Addgene 20672) respectively, and generated MIT-PP2A and MIG-RACK1 retrovirus in 293T cells by transient transfection.

Techniques: In Vitro, Reverse Transcription Polymerase Chain Reaction, Flow Cytometry, RNA Sequencing, Western Blot

Journal: eLife

Article Title: Natural Tr1-like cells do not confer long-term tolerogenic memory

doi: 10.7554/eLife.44821

Figure Lengend Snippet:

Article Snippet: Antibody , InVivoMAb mouse monoclonal anti-mouse Thy1.1/CD90.1 (clone 19E12) , BioXcell , Cat: BE0214 , 200 ug/mouse.

Techniques: Flow Cytometry

Journal: Experimental and Therapeutic Medicine

Article Title: Effect of Src tyrosine kinase on a rat model of asthma

doi: 10.3892/etm.2021.11095

Figure Lengend Snippet: Identification of rat bone marrow mesenchymal stem cells in vitro . CD45 and CD90 were labelled with Cy3 (red) and the cells were stained with DAPI (blue). The results were observed by fluorescence microscopy. (A) CD90 labelled with Cy3 fluorescence; (B) DAPI staining; (C) merged CD90 and DAPI staining. The positive rate of CD90 was >90%. (D) CD45 labelled with Cy3; (E) DAPI staining; (F) merged CD45 and DAPI staining. The positive rate of CD45 was <5%.

Article Snippet: Concentrated goat serum and Cy3-labeled Goat Anti-Rabbit IgG (cat. no. BA1034) were purchased from Wuhan Boster Biological Technology Co., Ltd.; Entranster TM - in vivo (cat. no. 18668-11-1) was purchased from Engreen Biosystem, Ltd.; Lipofectamine ® 2000 was purchased from Invitrogen (Thermo Fisher Scientific, Inc.); DNA marker was purchased from TransGen Biotech Co., Ltd.; RNAiso Plus, PrimeScript RT Reagent kit and SYBR Premix Ex Taq TM II (Tli RNaseH Plus) were purchased from Takara Bio, Inc.; IL-5 (cat. no. E-EL-R0558c; Elabscience Biotechnology, Inc.), IL-33 (cat. no. CSB-E14077r; Cusabio Technology LLC) and IFN-γ (cat. no. CSB-E04579r; Cusabio Technology LLC) ELISA kits, and CD90 (cat. no. E-AB-70323) and CD45 (cat. no. E-AB-16319) antibodies were purchased from Elabscience Biotechnology, Inc.

Techniques: In Vitro, Staining, Fluorescence, Microscopy