Oxidative Medicine and Cellular Longevity / 2016 / Article / Fig 3

Research Article

Cell Line-Dependent Variability of Coordinate Expression of p75NTR and CRABP1 and Modulation of Effects of Fenretinide on Neuroblastoma Cells

Figure 3

Metabolic viability and cell number of SH-EP1 cells transfected with CRABP1 siRNA or a scrambled control construct after treatment with fenretinide. (a) Alamar blue assay performed 60 h after treatment with fenretinide. Metabolic viability differs between CRABP1 knockdown (□) and scrambled control construct-transfected (■) cells () at 4, 10, and 13 μM fenretinide. Note that Alamar blue assay does not account for cells already lost to apoptosis at the time of assay. A representative Western blot shows CRABP1 and β-actin proteins in Wildtype (WT), scrambled construct-treated (SC), and CRABP1 siRNA-treated (siRNA) cells. The graph below the blot depicts the mean optical density and SEM for 3 blots performed. (b) Representative photomicrographs (10x) of untreated and fenretinide-treated (13 μM) CRABP1 knockdown, mock- (empty vector) transfected, and scrambled control construct-transfected SH-EP1 cells. As depicted in the graph, mock- and scrambled construct-treated cells showed greater cell loss (, Student’s -test; determinations) than CRABP1 knockdown cells after fenretinide treatment. The three cell lines demonstrated equivalent cell culture growth and survival under control conditions. (c) Alamar blue metabolic viability assay demonstrates equivalent cell culture growth and redox reserve for the three cell lines under control conditions (NS: no statistically significant difference).