c-series rat cytokine antibody array  (RayBiotech inc)

Journal: Frontiers in Cellular Neuroscience

Article Title: Redox and Calcium Alterations of a Müller Cell Line Exposed to Diabetic Retinopathy-Like Environment

doi: 10.3389/fncel.2022.862325

Figure Lengend Snippet: Effect of high glucose (HG) on rMC-1 cellular proliferation and cytokine-induced activation. (A) Scatter graph with bars represents the proliferation rate of rMC-1 cells, plated at a density of 60,000 cells/well and examined after 24 or 48 h. HG promoted rMC-1 cellular proliferation at 48 h, but not at 24 h. Absorbance values of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] are expressed as mean ± SEM of four independent experiments. Statistical significance is calculated by two-way ANOVA followed by Tukey’s post-hoc test (**** p < 0.0001). (B) Scatter graph with bars represents nitric oxide (NO) release from rMC-1 cells, indicative of cellular activation. rMC-1 cells were plated and maintained in culture for 24 h, then treated for 24 h with a cytokine mix (IL-1β, TNF-α, and IFN-γ). 48 h after plating, rMC-1 cells were lysed, and NO concentration was normalized over their total protein content. Absorbance values are expressed as mean ± SEM of five independent experiments. Statistical significance is calculated by two-way ANOVA followed by Tukey’s post-hoc test (** p < 0.005; *** p < 0.001; **** p < 0.0001). (C) Representative western blot image of inducible NO synthase (iNOS) expression of rMC-1 cells. iNOS is detectable only in HG-maintained rMC-1 cells treated with the cytokine mix. (D) Scatter graph with bars representing iNOS expression of untreated and CK-treated rMC-1 cells normalized over α-tubulin expression. Fold values are expressed as mean ± SEM of five independent experiments. Statistical significance is calculated by two-way ANOVA followed by Tukey’s post-hoc test (*** p < 0.001). NG, normal glucose; HG, high glucose; M, mannitol; CK, cytokines.

Article Snippet: The cytokine/chemokine profile analysis was performed according to manufacturer instructions (RayBio ® C-Series Rat Cytokine Antibody Array C1, RayBiotech Life, Inc., Georgia, USA); to assess a possible cross-reaction with the human or mouse cytokines used to promote the activation process, a sample of medium spiked with the CK mix was analyzed and the obtained value was subtracted from experimental data.

Techniques: Activation Assay, Concentration Assay, Western Blot, Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Redox and Calcium Alterations of a Müller Cell Line Exposed to Diabetic Retinopathy-Like Environment

doi: 10.3389/fncel.2022.862325

Figure Lengend Snippet: Effect of HG on cytokine and chemokine release profile in CK-activated rMC-1 cells. Bar plot represents cytokines and chemokines profile of rMC-1 cells exposed to HG and eventually treated with the CK mix (HG CK); the results are shown as values obtained from HG CK condition normalized over HG. Fold values are expressed as mean ± SEM of three independent experiments.

Article Snippet: The cytokine/chemokine profile analysis was performed according to manufacturer instructions (RayBio ® C-Series Rat Cytokine Antibody Array C1, RayBiotech Life, Inc., Georgia, USA); to assess a possible cross-reaction with the human or mouse cytokines used to promote the activation process, a sample of medium spiked with the CK mix was analyzed and the obtained value was subtracted from experimental data.

Techniques: