Deficiency of CCAR2 or Hsp60 reduces expression of survivin mRNA. SH-SY5Y (A–E) or BE(2)-M17 cells (E) were transfected with Universal (siU), CCAR2 (siC), or Hsp60 (siH) siRNA. (A,B) Forty-eight hours later, the level of survivin protein was examined by western blotting. Cells deficient in CCAR2 and Hsp60 were treated with 25 μM MG132 (A) or 100 μM chloroquine (B) 4 h or 24 h prior to cell lysis, respectively. (C,D) The level of survivin mRNA in each group of siRNA-transfected cells was measured by RT-PCR. (C) Levels were normalized against β-actin and quantified using ImageJ software. The relative level of survivin mRNA is expressed as the mean ± standard error of the mean (SEM) (n = 3). Asterisks (*) denote statistically significant differences (p < 0.05, one-way ANOVA). (D) Two different siRNAs targeting CCAR2 and Hsp60 were used to knock down their expressions. (E) The level of each protein was examined by western blotting. The relative level is expressed as the mean ± standard error of the mean (SEM) (n = 3). Indicators (*, #) denote statistically significant differences from the corresponding control cells (p < 0.05, one-way ANOVA).