Journal: Matrix Biology Plus

Article Title: The importance of matrix in cardiomyogenesis: Defined substrates for maturation and chamber specificity

doi: 10.1016/j.mbplus.2024.100160

Figure Lengend Snippet: ECMP Plate Array A) Layout of arrayed ECMPs in a 96 well plate. hESC are seeded at 1.24 x 10 4 c e l l s / cm 2 to allow proliferation over 5 days. B) Images of hESC using Olympus confluency checker where green donates culture plastic and purple denotes cells. Images taken on day 5 at 10X magnification, Scale bar 500 μ M . C) Graphical comparison of the viability and confluency of all 63 ECMP combinations to highlight similarities and differences of the ECMP combinations on the hESC. D) List of protein combinations used to generate the ECMP array with the proteins name abbreviation, the location of the well in the 96 well plate and the well plate number referenced to the X axis of the comparison of confluency and viability graph.

Article Snippet: After seeding hESCs across the array, we measured confluency on each ECMP combination over 5 days using the Olympus Confluency Checker.

Techniques: Comparison

Journal: Diabetes, obesity & metabolism

Article Title: Preclinical evaluation of tyrosine kinase 2 inhibitors for human beta-cell protection in type 1 diabetes

doi: 10.1111/dom.14104

Figure Lengend Snippet: TYK2 inhibition prevents IFNα-induced STAT1/2 phosphorylation and HLA-ABC, CXCL10, MX1 and CHOP upregulation in human islet cells. Dispersed human islets were left untreated (CTL) or pretreated with 1 μM of TYK2 inhibitor (iA and iB for TYK2iA and B, respectively) for 2 hours. Then IFNα (2000 U/mL) was added for 24 hours in the continuous presence of the inhibitors. (A) Protein expression was measured by western blot and representative images of six independent experiments are shown. Densitometry results are shown for pSTAT1 (B) and pSTAT2 (C) and the values were normalized by β-actin. The mRNA expression of HLA-ABC (D), CXCL10 (E), MX1 (F) and CHOP (G) was analysed by RT-qPCR and the values were normalized by β-actin. (H) CXCL10 protein secretion in the supernatant fraction was determined by ELISA. In all experiments, the values were normalized by cells treated with IFNα without TYK2 inhibitors, considered as 1. Results are mean ± SEM of six (A-G) or five (H) independent experiments. *P < .05, **P < .01 and ***P < .001 vs. control (CTL NT); †P < .05, ††P < .01 and †††P < .001 vs. CTL IFNα; one-way ANOVA. (I) ICC of MHC class I (red), insulin (green) and Hoechst (blue) in human islets treated with IFNα with or without TYK2 inhibitors as described above (magnification ×40)

Article Snippet: A series of in vitro studies investigated the functional consequences of the binding of the TYK2 inhibitors to the TYK2 JH2 pseudokinase domain in isolated human peripheral blood mononuclear cells (PBMCs; Confluence Discovery Technologies, St. Louis, MO, USA).

Techniques: Inhibition, Expressing, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Diabetes, obesity & metabolism

Article Title: Preclinical evaluation of tyrosine kinase 2 inhibitors for human beta-cell protection in type 1 diabetes

doi: 10.1111/dom.14104

Figure Lengend Snippet: TYK2 inhibitors revert IFNα-induced CXCL10 and MX1 but not HLA-ABC and CHOP expression in EndoC-βH1 cells. EndoC-βH1 cells were pretreated for 24 hours with IFNα (2000 U/mL). The TYK2 inhibitors (iA and iB for TYK2iA and B, respectively, 1 μM) were then added for an additional 24 hours in the continuous presence of IFNα. The mRNA expression of CXCL10 (A), MX1 (B), HLA-ABC (C) and CHOP (D) was analysed by RT-qPCR and the values were normalized by β-actin and then by cells treated with IFNα without TYK2 inhibitors, considered as 1. Results are mean ± SEM of six independent experiments. *P < .05, **P < .01 and ***P < .001 vs. control (CTL NT); †††P < .001 vs. CTL IFNα; one-way ANOVA

Article Snippet: A series of in vitro studies investigated the functional consequences of the binding of the TYK2 inhibitors to the TYK2 JH2 pseudokinase domain in isolated human peripheral blood mononuclear cells (PBMCs; Confluence Discovery Technologies, St. Louis, MO, USA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Diabetes, obesity & metabolism

Article Title: Preclinical evaluation of tyrosine kinase 2 inhibitors for human beta-cell protection in type 1 diabetes

doi: 10.1111/dom.14104

Figure Lengend Snippet: TYK2 inhibition prevents apoptosis and the expression of inflammatory and ER stress markers induced by IFNα + IL-1β in dispersed human islets. Dispersed human islets were left untreated (CTL) or pretreated with 1 μM of TYK2 inhibitor (iA and iB for TYK2iA and B, respectively) for 2 hours. Afterwards, IFNα (2000 U/mL) + IL-1β (50 U/mL) were added for 24 hours in the continuous presence of the inhibitors. (A) Apoptosis was evaluated using HO/PI staining. (B, K) Protein expression was measured by western blot and representative images of three (K) or five to eight (B) independent experiments are shown. Densitometry results normalized by β-actin are shown for pSTAT1 (C), pSTAT2 (D) and ATF3 (L). The mRNA expression of HLA-ABC (E), CXCL10 (F), MX1 (H), CHOP (I) and ATF3 (J) was analysed by RT-qPCR and the values were normalized by β-actin. (G) CXCL10 protein secretion in the supernatant fraction was determined by ELISA. In all experiments, values were normalized by cells treated with IFNα + IL-1β without TYK2 inhibitors, considered as 1. Results are mean ± SEM of three to nine independent experiments. *P < .05, **P < .01 and ***P < .001 vs. control (CTL NT); †P < .05, ††P < .01 and †††P < .001 vs. CTL IFNα + IL-1β; one-way ANOVA

Article Snippet: A series of in vitro studies investigated the functional consequences of the binding of the TYK2 inhibitors to the TYK2 JH2 pseudokinase domain in isolated human peripheral blood mononuclear cells (PBMCs; Confluence Discovery Technologies, St. Louis, MO, USA).

Techniques: Inhibition, Expressing, Staining, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Diabetes, obesity & metabolism

Article Title: Preclinical evaluation of tyrosine kinase 2 inhibitors for human beta-cell protection in type 1 diabetes

doi: 10.1111/dom.14104

Figure Lengend Snippet: TYK2 inhibitors do not sensitize human beta cells to viral infection. EndoC-βH1 cells (A, C, E, G) and dispersed human islets (B, D, F, H) were left untreated (CTL) or pretreated with 1 μM of TYK2 inhibitor (iA and iB for TYK2iA and B, respectively) for 2 hours. Afterwards, cells were infected or not (MOCK) with CVB1 (MOI 0.1) or CVB5 (MOI 5) for 24 hours in the continuous presence of the inhibitors. (A, B) Apoptosis was evaluated using HO/PI staining. (C, D) Viral replication was quantified by viral titration and presented as relative viral titer compared with the control condition (CTL) for each virus. (E, F) The expression of the viral capsid protein (VP1) was evaluated by western blot and representative images of four (E) and three (F) independent experiments are shown. Densitometry results are shown (G, H) and the values were normalized by α-tubulin and then by each control condition (without TYK2 inhibitor), considered as 1. Results are mean ± SEM of four (EndoC-βH1 cells) and three (dispersed human islets) independent experiments. *P < .05 and **P < .01 vs. CTL MOCK; one-way ANOVA

Article Snippet: A series of in vitro studies investigated the functional consequences of the binding of the TYK2 inhibitors to the TYK2 JH2 pseudokinase domain in isolated human peripheral blood mononuclear cells (PBMCs; Confluence Discovery Technologies, St. Louis, MO, USA).

Techniques: Infection, Staining, Titration, Expressing, Western Blot