bio-plex pro™ human th17 cytokine 15-plex panel  (Bio-Rad)

Journal: Human Molecular Genetics

Article Title: Novel mutations in the WFS1 gene are associated with Wolfram syndrome and systemic inflammation

doi: 10.1093/hmg/ddab040

Figure Lengend Snippet: Proinflammatory cytokines production by WS PBMCs. ( A ) Schematic heat map representation of Bio-Plex Pro Human Th17 cytokine panel. The cytokine profile was analyzed in supernatants from PBMCs from healthy donors (Ctrl) and from proband’s PBMCs (WS). The contents of selected cytokines were also analyzed by single-cytokine ELISA test and data were reported in ( B ). ( C ) Real-time PCR analysis of TNF-α, IL-1β, IL-6 transcripts in freshly purified PBMCs from healthy controls (Ctrl) or the WS patient. Data were normalized to the expression of ACTB (encoding β-actin) and presented relative to results in healthy controls untreated cells (dotted line, 1-fold). In ( B ) and ( C ) data were analyzed by two-tailed Student’s t -test. * P < 0.05; * * P < 0.01; * * * P < 0.001; * * * * P < 0.0001. ( D ) Schematic heat map representation of Cytokine 25-Plex Human ProcartaPlex™ Panel 1B. The cytokine profile was analyzed in sera from healthy donors (Ctrl), type 1 diabetes subjects (T1D) and the WS proband.

Article Snippet: The cytokine profile was analyzed in cell culture supernatants by using a 15-plex immunoassay (Bio-Plex Pro Human Th17 cytokine panel; Bio-Rad, CA, USA) or in sera by using a Cytokine 25-Plex Human ProcartaPlex™ Panel 1B (Invitrogen) and a MAGPIX system (Luminex Corporation).

Techniques: Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Purification, Expressing, Two Tailed Test

Journal: Human Molecular Genetics

Article Title: Novel mutations in the WFS1 gene are associated with Wolfram syndrome and systemic inflammation

doi: 10.1093/hmg/ddab040

Figure Lengend Snippet: Production of proinflammatory cytokines after silencing of the WFS1 gene in PBMCs from healthy donors. ( A ) The mRNA level of WFS1 after transfection with WFS1 siRNA was detected by real-time PCR in PBMCs of three healthy donors. Control cells were treated with vehicle alone (DOTAP). Data were normalized to the expression of ACTB (encoding β-actin) and presented relative to results in untreated cells (dotted line, 1-fold). One experiment is shown representative of three. Data were analyzed by one-way ANOVA followed by post hoc Bonferroni’s test. ( B ) Schematic heat map representation of Bio-Plex Pro Human Th17 cytokine panel. The cytokine profile was analyzed in supernatants from PBMCs of healthy donors untransfected or transfected with siWFS1 or a scramble siRNA. Vehicle alone (DOTAP) was used as a control. Individual expression levels of selected cytokines were reported in ( C ). Data were analyzed by one-way ANOVA followed by post hoc Bonferroni’s test. * * P < 0.01; * * * P < 0.001; * * * * P < 0.0001.

Article Snippet: The cytokine profile was analyzed in cell culture supernatants by using a 15-plex immunoassay (Bio-Plex Pro Human Th17 cytokine panel; Bio-Rad, CA, USA) or in sera by using a Cytokine 25-Plex Human ProcartaPlex™ Panel 1B (Invitrogen) and a MAGPIX system (Luminex Corporation).

Techniques: Transfection, Real-time Polymerase Chain Reaction, Expressing