Figure 4. Knockdown of peroxiredoxin-3 (PRDX3)-induced oxidative stress and mitochondrial dysfunction.

HTR8 cells were transiently transfected with PRDX3 shRNA plasmid (sh-PRDX3) or scramble shRNA (sh-scr). (A) The messenger RNA expressions of PRDX family members (PRDX1-6) were determined by qRT-PCR with normalisation to 18s rRNA level. (B) Reactive oxygen species production, (C) mitochondrial membrane potential (ΔΨm) and (D) adenosine triphosphate concentration were assayed at 36 hours post-transfection. Results for reactive oxygen species and mitochondrial membrane potential were expressed as arbitrary units, whereas adenosine triphosphate (ATP) level was expressed as nmol/mg protein. (E,F) Twenty-four hours after transfection, HTR8 cells were treated with 100 μM cholic acid, deoxycholic acid and chenodeoxycholic acid for 24 hours. (E) Cell viability assay and (F) quantification of lactate dehydrogenase release were applied. The results were expressed as percentage of control. Data were shown as the mean ± standard error of mean. Statistical significance was determined by two-tailed Student’s t test. (**p < 0.01, ***p < 0.001 versus the sh-scr group).