Journal: PLoS ONE

Article Title: COUP-TFII Controls Mouse Pancreatic β-Cell Mass through GLP-1-β-Catenin Signaling Pathways

doi: 10.1371/journal.pone.0030847

Figure Lengend Snippet: In pancreatic sections of 5-day-old mice (A) β-cell proliferation was assessed by double staining insulin/Ki67 (B) β-cell neogenesis activation was evaluated through quantification of duct-associated insulin+ β-cells per unit of total tissue area. 832/13 INS-1 β-cells were electroporated with scrambled or specific COUP-TFII siRNA and cultured in INS-1 medium for 24 h or 48 h. (C) Relative COUP-TFII mRNA levels determined by RT-qPCR at 48 h. (D) Effect of COUP-TFII siRNA on cell viability analyzed by MTT assay (n = 5 electroporations). (E) Effect of COUP-TFII siRNA on β-cell proliferation. 832/13 INS-1 β-cells were cultured in INS-1 medium for 48 h and stained with BrdU. Cells were analyzed by flow cytometry (n = 4 electroprations). (F) Effect of COUP-TFII siRNA on β-cells apoptosis and comparison of the apoptotic rate with cell treated with a rat cytokine mix containing 25 ng/ml TNF-α, 10 ng/ml IL-1β and 10 ng/ml INF-γ during 24 h. 832/13 INS-1 β-cells were electroporated with scrambled or COUP-TFII siRNA and 48 h later they were stained with DiOC6(3) and analyzed by flow cytometry (n = 4 electroporations). * Significant difference between values linked by brackets at P <0.03.

Article Snippet: To induce apoptosis, cells were treated with a rat cytokine mix containing 25 ng/ml TNF-α (catalogue number 400-14, Peprotech), 10 ng/ml IL-1β (catalogue number 400-01, Peprotech), and 10 ng/ml INF-γ (catalogue number 400-20, Peprotech) during 24 h.

Techniques: Double Staining, Activation Assay, Cell Culture, Quantitative RT-PCR, MTT Assay, Staining, Flow Cytometry, Comparison

Journal: Experimental Biology and Medicine

Article Title: In vitro psoriasis models with focus on reconstructed skin models as promising tools in psoriasis research

doi: 10.1177/1535370217710637

Figure Lengend Snippet: Overview of the four different psoriatic skin equivalents created by cytokine stimulation of healthy skin equivalents, except for the self-assembly model. Macroscopic images are showing the different matrix types followed by histological pictures or stainings of differentiation markers such as filaggrin in the synthetic membrane model (A)41 and CK16 in the DED-based model (C)61 before and after stimulation with indicated cytokines. In the self-assembly model (B), psoriatic fibroblasts and psoriatic keratinocytes were used to generate a psoriatic full-thickness (FT) 3D skin model.78 Addition of cytokines to this model compensated for the lack of immune cells.94 Psoriatic fibroblasts, but healthy KCs, were used by MatTek to establish their collagen-based FT 3D psoriatic model (D). Finally, proteins and genes known to be differentially expressed in the psoriatic skin models versus healthy controls (except in B, deregulated genes are described after comparison of psoriatic substitutes treated or not with cytokines) are summarized. Models B to D represent FT models, whereas model A contains only a differentiated epidermal part. CK16: Keratin 16; DED: de-epidermized dermis. Scale bar: 100 µm. (Histological images and image of self-assembly model (see referred papers) are reprinted with permission from Elsevier and Future Medicine Ltd)

Article Snippet: • MatTek FT , NHEK + PHDF , MatTek cytokine mix , Drug testing: PTTC, calcipotriol, IL-4, coal tar, psoriasin gel, delphinidin , 84 – 88.

Techniques:

Journal: Experimental Biology and Medicine

Article Title: In vitro psoriasis models with focus on reconstructed skin models as promising tools in psoriasis research

doi: 10.1177/1535370217710637

Figure Lengend Snippet: Commercially available psoriasis 3D skin models for in vitro applications

Article Snippet: • MatTek FT , NHEK + PHDF , MatTek cytokine mix , Drug testing: PTTC, calcipotriol, IL-4, coal tar, psoriasin gel, delphinidin , 84 – 88.

Techniques: In Vitro, Ex Vivo

Journal: Experimental Biology and Medicine

Article Title: In vitro psoriasis models with focus on reconstructed skin models as promising tools in psoriasis research

doi: 10.1177/1535370217710637

Figure Lengend Snippet: Overview of applications using psoriasis(-induced) skin substitutes

Article Snippet: • MatTek FT , NHEK + PHDF , MatTek cytokine mix , Drug testing: PTTC, calcipotriol, IL-4, coal tar, psoriasin gel, delphinidin , 84 – 88.

Techniques: Expressing, Migration, In Vivo

Journal: Experimental Biology and Medicine

Article Title: In vitro psoriasis models with focus on reconstructed skin models as promising tools in psoriasis research

doi: 10.1177/1535370217710637

Figure Lengend Snippet: Overview of the different types of psoriasis models with pros and cons, and their most current applications

Article Snippet: • MatTek FT , NHEK + PHDF , MatTek cytokine mix , Drug testing: PTTC, calcipotriol, IL-4, coal tar, psoriasin gel, delphinidin , 84 – 88.

Techniques: Ex Vivo, Migration, Activity Assay, Transgenic Assay

Journal: PLoS ONE

Article Title: Epstein Barr virus Latent Membrane Protein-1 enhances dendritic cell therapy lymph node migration, activation, and IL-12 secretion

doi: 10.1371/journal.pone.0184915

Figure Lengend Snippet: Human monocytes were isolated from buffy coat and cultured in the presence of GM-CSF and IL-4 for 5 days to generate DC. On day 5, cells were infected with Ad5-LMP1 or Ad5-GFP at an MOI of 100 in the presence of doxycycline. Mimic cytokine maturation cocktail was added on day 5 as a positive control. (A) Cells were analyzed by flow cytometry and representative forward vs. side scatter plots are displayed. (B) Images of DC were taken on day 7 before harvesting for maturation staining. Scale bar equals 100μm. (C) Representative gating strategy used to determine maturation of MDDC. (D) DC were stained for maturation markers, and analyzed by flow cytometry. (E) Supernatant was collected every 12 hours for 2 days and cytokine measured by CBA. TNF-α, IL-6 and IL-1β levels for Mimic sample may represent residual protein from the Mimic cocktail. Data represent results from three independent experiments.

Article Snippet: As a positive control, cytokine mix Mimic (5 ng/ml TNF-α (Peprotech), 5ng/ml IL-1b (Peprotech), 750ng/ml IL-6 (Peprotech), and 1ug/ml PGE2 (Sigma)) was used to mature DC.

Techniques: Isolation, Cell Culture, Infection, Positive Control, Flow Cytometry, Staining