Journal: PLoS ONE

Article Title: Folate Deficiency Triggers an Oxidative-Nitrosative Stress-Mediated Apoptotic Cell Death and Impedes Insulin Biosynthesis in RINm5F Pancreatic Islet β–Cells: Relevant to the Pathogenesis of Diabetes

doi: 10.1371/journal.pone.0077931

Figure Lengend Snippet: (A) Production of intracellular ROS in the designated cells cultivated under either FC or FD condition was evaluated flowcytometrically using DCF-DA as the probe. At the end of 2-week cultivation period under FD condition, a nearly 3-fold increase in ROS production, as reflected by the DCF fluorescence intensity, could be detected as compared to FC control. The values shown are mean ± standard errors (n = 3 of individual experiments). *p<0.05 vs. control. (B) The production of homocysteine (Hcy) was followed using an enzyme-assisted spectrophotometric detection of H 2 S production which is directly proportional to the concentration of Hcy. The results were expressed as µM Hcy/10 6 cells. Each bar represents an average of triplicate determinations. *p<0.05 v.s control. (C) Direct measurement of H 2 O 2 released by RINm5F cells using Amplex red hydrogen peroxide assay kit. The results were expressed as µM H 2 O 2 /10 5 cells. It is apparent that H 2 O 2 production by RINm5F cells cultivated under FD condition was higher than FC control. *p<0.05 v.s control. (D) The production of OH • in either FC or FD condition of RINm5F cells was determined flowcytometrically using hydroxyphenyl fluorescein (HPF) as a probe. *p<0.05 v.s control.

Article Snippet: The homocysteine levels of the culture medium were then analyzed by an automated autoanalyzer [Hitachi 917 (7170)] based on the total homocysteine biochemical assay kit developed by Formosa Biomedical Technology Co.

Techniques: Fluorescence, Control, Concentration Assay, Amplex Red Hydrogen Peroxide Assay