Journal: Cells

Article Title: FGF2 Inhibits Early Pancreatic Lineage Specification during Differentiation of Human Embryonic Stem Cells

doi: 10.3390/cells9091927

Figure Lengend Snippet: Pancreatic differentiation in presence of growth factors. ( a ) Gene expression of PDX1 , HLXB9 , HNF1B , HNF6 , SHH, TBX1, SOX9, AFP, and CDX2 measured by RT-qPCR in human embryonic stem cells (hESC)s at d0, d4, and d8 of differentiation treated ± 5 or 50 ng/mL fibroblast growth factor (FGF)2, FGF7, FGF10, or epidermal growth factor (EGF) in stage 2 medium. Values are means ± SEM, n = 4–6. 5/50 = 5 or 50 ng/mL growth factor, C = control medium without growth factors. ( b ) Fluorescence micrographs after pancreatic differentiation illustrating the protein expression of PDX1/HNF1B after a 4-day treatment ± 5 or 50 ng/mL FGF2, FGF7, FGF10, and EGF. Nuclei were counterstained with DAPI. Scale bar = 200 µm and 20 µm for higher magnification of the control. ( c ) Quantification of cell expansion using ± 50 ng/mL growth factor. Data are presented as mean (log2) ± SEM, n = 3. ( d ) Quantification of PDX1-positive cells upon treatment ± 5 or 50 ng/mL FGF2, FGF7, FGF10, and EGF at d8. Percentages are expressed as means ± SEM. *** = p ≤ 0.001, ** = p ≤ 0.01 compared to control, ANOVA plus Dunnett’s post-test.

Article Snippet: The primary antibodies anti-HNF1B (SantaCruz, sc-22840, Santa Cruz, CA, USA) and anti-PDX1 (R&D systems, AF2419, Minneapolis, MN, USA) were used.

Techniques: Gene Expression, Quantitative RT-PCR, Control, Fluorescence, Expressing

Journal: Cells

Article Title: FGF2 Inhibits Early Pancreatic Lineage Specification during Differentiation of Human Embryonic Stem Cells

doi: 10.3390/cells9091927

Figure Lengend Snippet: Gene expression changes in response to different FGF2 concentrations. ( a ) hESCs were cultured in the presence of 5–50 ng/mL FGF2 during differentiation into PDX1-positive cells and compared to a control condition (=C). Gene expression changes of PDX1 , HNF1B , SHH, and TBX1 are presented as means ± SEM, n = 5–12. *** = p ≤ 0.001, ** = p ≤ 0.01, * = p ≤ 0.05 compared to control, ANOVA plus Dunnett’s post-test. ( b ) Effect of SHH inhibition by 10 µM Gant 58 or 2.5 µM Sant1 in stage 2 medium ± 50 ng/mL FGF2. Gene expression changes of PDX1 and SHH are presented as means ± SEM, n = 4–5. *** = p ≤ 0.001, compared to control, ANOVA plus Dunnett’s post-test.

Article Snippet: The primary antibodies anti-HNF1B (SantaCruz, sc-22840, Santa Cruz, CA, USA) and anti-PDX1 (R&D systems, AF2419, Minneapolis, MN, USA) were used.

Techniques: Gene Expression, Cell Culture, Control, Inhibition

Journal: Cells

Article Title: FGF2 Inhibits Early Pancreatic Lineage Specification during Differentiation of Human Embryonic Stem Cells

doi: 10.3390/cells9091927

Figure Lengend Snippet: Inhibition of FGFR1c/3c rescues pancreatic development in presence of FGF2 and FGF7. ( a ) Gene expression changes of PDX1 , HNF1B , SHH , and TBX1 in d8 cells treated ± 10 ng/mL FGF2 (F2) and the FGFR1c/3c small molecule inhibitor PD-173074 (PD) with a concentration of 100 nM (PD). Data are means ± SEM, n = 4–5. *** = p ≤ 0.001, ** = p ≤ 0.01, * p = ≤0.05 ANOVA plus Tukey’s post-test, F2 treatment compared to F2 + PD and PD only. ( b ) Representative immunofluorescence staining of PDX1 in d8 control cells, cells treated with F2 and F2 + PD. Nuclei were counterstained with DAPI. Scale bar = 100 µm. ( c ) Gene expression changes of PDX1 , SOX9 , AFP , and CDX2 in d8 cells treated ± 50 ng/mL FGF7 (F7) and ± PD-173074 with a concentration of 10 or 100 nM (10, 100). Values are means ± SEM, n = 3–6, * = p ≤ 0.05, ANOVA plus Tukey’s post-test. ( d ) Quantification of PDX1-positive cells at d8 upon treatment ± 50 ng/mL FGF7 and ± PD-173074 with a concentration of 10 or 100 nM. Percentages are expressed as means ± SEM. * = p ≤ 0.05, ANOVA plus Tukey’s post-test.

Article Snippet: The primary antibodies anti-HNF1B (SantaCruz, sc-22840, Santa Cruz, CA, USA) and anti-PDX1 (R&D systems, AF2419, Minneapolis, MN, USA) were used.

Techniques: Inhibition, Gene Expression, Concentration Assay, Immunofluorescence, Staining, Control

Journal: Viruses

Article Title: Efficient Sensing of Avian Influenza Viruses by Porcine Plasmacytoid Dendritic Cells

doi: 10.3390/v3040312

Figure Lengend Snippet: List of influenza A virus (IAV) isolates used in this study.

Article Snippet: HPAIV A/Turkey/Turkey/05, A/Cygnus/Italy/742/06, A/Mallard/Italy/835/06 (H5N1), A/Turkey/Italy/4580/98, A/Ostrich/Italy/2332/00 as well as LPAIV A/Turkey/Italy/3675/99 (H7N1) were kindly provided by Dr. W. Dundon, (IZSV Istituto Zooprofilattico Sperimentale delle Venezie, Venice, Italy), human isolates A/New Caledonia/20/99 (H1N1) A/Wisconsin/67/05 (H3N2) were kindly given by Drs. W. Wunderli and Y. Thomas (National Influenza Reference Center, Geneva University Hospital, Switzerland), swine IAV A/Swine/Belgium/1/98 and A/Swine/Flanders/1/98 (H1N1) were kindly received from Dr. K. Van Reeth (Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Belgium) and A/Hong Kong/8/68 (H3N2) and A/Swine/Iowa/1976/31 (H1N1) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Virus

Journal: Viruses

Article Title: Efficient Sensing of Avian Influenza Viruses by Porcine Plasmacytoid Dendritic Cells

doi: 10.3390/v3040312

Figure Lengend Snippet: Relation between virus dose, pDC percentage and IFN-α response. Porcine pDC were stimulated with H5N1 TT05, H7N1 TI99, H1N1 SI76 and H1N1 NC99 (listed in ) at 100, 1 or 0.01 HAU per well or left non-infected. ( A ) CD4/CD172a plots are shown for H5N1 TT05 infection at the tested doses and for non-infected cells to illustrate the gate for pDC definition. Numbers in the plots indicate the pDC percentage present in the small square. ( B ) pDC percentage after stimulation with different virus doses. ( C ) IFN-α secretion in the supernatant in function of the virus dose.

Article Snippet: HPAIV A/Turkey/Turkey/05, A/Cygnus/Italy/742/06, A/Mallard/Italy/835/06 (H5N1), A/Turkey/Italy/4580/98, A/Ostrich/Italy/2332/00 as well as LPAIV A/Turkey/Italy/3675/99 (H7N1) were kindly provided by Dr. W. Dundon, (IZSV Istituto Zooprofilattico Sperimentale delle Venezie, Venice, Italy), human isolates A/New Caledonia/20/99 (H1N1) A/Wisconsin/67/05 (H3N2) were kindly given by Drs. W. Wunderli and Y. Thomas (National Influenza Reference Center, Geneva University Hospital, Switzerland), swine IAV A/Swine/Belgium/1/98 and A/Swine/Flanders/1/98 (H1N1) were kindly received from Dr. K. Van Reeth (Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Belgium) and A/Hong Kong/8/68 (H3N2) and A/Swine/Iowa/1976/31 (H1N1) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Virus, Infection

Journal: Viruses

Article Title: Efficient Sensing of Avian Influenza Viruses by Porcine Plasmacytoid Dendritic Cells

doi: 10.3390/v3040312

Figure Lengend Snippet: List of reverse genetic viruses used in this study.

Article Snippet: HPAIV A/Turkey/Turkey/05, A/Cygnus/Italy/742/06, A/Mallard/Italy/835/06 (H5N1), A/Turkey/Italy/4580/98, A/Ostrich/Italy/2332/00 as well as LPAIV A/Turkey/Italy/3675/99 (H7N1) were kindly provided by Dr. W. Dundon, (IZSV Istituto Zooprofilattico Sperimentale delle Venezie, Venice, Italy), human isolates A/New Caledonia/20/99 (H1N1) A/Wisconsin/67/05 (H3N2) were kindly given by Drs. W. Wunderli and Y. Thomas (National Influenza Reference Center, Geneva University Hospital, Switzerland), swine IAV A/Swine/Belgium/1/98 and A/Swine/Flanders/1/98 (H1N1) were kindly received from Dr. K. Van Reeth (Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Belgium) and A/Hong Kong/8/68 (H3N2) and A/Swine/Iowa/1976/31 (H1N1) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Binding Assay