Journal: Cell metabolism

Article Title: Comprehensive Proteomics Analysis of Stressed Human Islets Identifies GDF15 as a Target for Type 1 Diabetes Intervention

doi: 10.1016/j.cmet.2019.12.005

Figure Lengend Snippet: (A) GDF15 and SPP1 levels in human islets treated with the combination of cytokines IL-1β+IFN-γ were measured by label-free proteomics, western blot, and qPCR. The western blot image is shown in Figure S2.

Article Snippet: To study the protective effect of GDF15 and SPP1, we pretreated EndoC βH1 cells and human islets with human recombinant 100 ng/mL GDF15 (Sino Biologicals) and 50 ng/mL SPP1 (GenTex), followed by cytokine treatment for 24 h. Recombinant GDF15 was previously tested for TGF-β contamination by ELISA, confirming absence of TGF-β in the sample.

Techniques: Western Blot

Journal: Cell metabolism

Article Title: Comprehensive Proteomics Analysis of Stressed Human Islets Identifies GDF15 as a Target for Type 1 Diabetes Intervention

doi: 10.1016/j.cmet.2019.12.005

Figure Lengend Snippet: (A) Network analysis using MetaCore of downstream signaling of GDF15 and SPP1. Note that both GDF15 and SPP1 regulate the apoptotic pathway by different but overlapping signaling.

Article Snippet: To study the protective effect of GDF15 and SPP1, we pretreated EndoC βH1 cells and human islets with human recombinant 100 ng/mL GDF15 (Sino Biologicals) and 50 ng/mL SPP1 (GenTex), followed by cytokine treatment for 24 h. Recombinant GDF15 was previously tested for TGF-β contamination by ELISA, confirming absence of TGF-β in the sample.

Techniques:

Journal: Cell metabolism

Article Title: Comprehensive Proteomics Analysis of Stressed Human Islets Identifies GDF15 as a Target for Type 1 Diabetes Intervention

doi: 10.1016/j.cmet.2019.12.005

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: To study the protective effect of GDF15 and SPP1, we pretreated EndoC βH1 cells and human islets with human recombinant 100 ng/mL GDF15 (Sino Biologicals) and 50 ng/mL SPP1 (GenTex), followed by cytokine treatment for 24 h. Recombinant GDF15 was previously tested for TGF-β contamination by ELISA, confirming absence of TGF-β in the sample.

Techniques: Plasmid Preparation, Recombinant, Modification, Produced, Enzyme-linked Immunosorbent Assay, LAL Assay, Caspase-Glo Assay, Expressing, Software

Journal: Cancers

Article Title: MiR-181 Family Modulates Osteopontin in Glioblastoma Multiforme

doi: 10.3390/cancers12123813

Figure Lengend Snippet: MiR-181 family regulates OPN. ( A ) Schema showing the derivation of M0 and M2 polarized macrophages that are the dominant phenotype of macrophages in gliomas. ( B ) Transient transfection of M0 or M2 macrophages on day 6 for 48 h with either non-targeting control (NC) or miR-181a/b/c/d-5p mimics. MiR overexpression levels were confirmed by qRT-PCR. ( C ) Analysis of the resulting cells revealed that overexpression of miR-181a/b/c/d mimics resulted in a decrease in OPN gene expression at the RNA level (qRT-PCR) and ( D ) the protein level (ELISA). Data were normalized against the levels of U6 small RNA for all qRT-PCR experiments. The data represent the mean ± SD ( n = 3) in three independent experiments. Data is cumulative of 3 independent experiments run in triplicates. Statistical significance was determined using an unpaired Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: OPN levels in the supernatant were measured by enzyme-linked immunosorbent assay (ELISA) 48 h after transfection using the OPN duo set ELISA (DY1433, R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Transfection, Control, Over Expression, Quantitative RT-PCR, Gene Expression, Enzyme-linked Immunosorbent Assay