The effects of aminochrome on β-tubulin (a–d) and the accumulation of autophagosomes in the cytosol of cells treated with aminochrome (e). RCSN-3 cells were treated with cell culture medium (a), 100 μM dicoumarol (b), 50 μM aminochrome (c), and 50 μM aminochrome and 100 μM dicoumarol (d) and incubated for 48 h as described by Paris et al. 2010 [15]. In (e) the number of autophagosomes per cell was determined by incubating RCSN3 cells for 24 h with cell culture medium (Co), 100 μM dicoumarol (Di), 10 μM vinblastine (V), 20 μM aminochrome (AM), 20 μM aminochrome and 100 μM dicoumarol (AMDi), 20 μM aminochrome and 100 μM dicoumarol and 10 μM vinblastine (AMDiV). As a positive control, cells were incubated with cell culture medium without bovine serum (Co+). The statistical significance was assessed using ANOVA for multiple comparisons and Student’s t test (***). This experiment was performed as described by Paris et al. 2011 [40].
