Fig. 5

Silencing of PNPLA8 increases cellular reactive oxygen species levels and decreases cell viability and cell migration. A Real-time PCR showing mRNA levels of PNPLA8 in SUM159PT and Hs578T cell lines treated with PNPLA8 siRNA or scrambled siRNA. Data are represented as mean values ± S.D. of 3 independent experiments. B Protein expression levels of PNPLA8 in SUM159PT and Hs578T cell lines treated with PNPLA8 siRNA or scrambled siRNA. GAPDH was used as a control to confirm equal loading of protein. C Flow cytometry analysis of mitochondrial reactive oxygen species (ROS) using MitoSOX in SUM159PT and Hs578T cell lines treated with PNPLA8 siRNA or scrambled siRNA as controls. Values represent MitoSOX intensity mean values ± S.D. of 3 independent experiments. D Flow cytometry analysis of reactive oxygen species (ROS) using DCFDA in SUM159PT and Hs578T cell lines treated with PNPLA8 siRNA or scrambled siRNA as controls. Values represent DCFDA intensity mean values ± S.D. of 3 independent experiments. E Representative images of migrated cells of SUM159PT and Hs578T cell lines treated with PNPLA8 siRNA or scrambled siRNA as control. F Number of migrated cells of SUM159PT and Hs578T cell lines treated with PNPLA8 siRNA or scrambled siRNA as control. Migrated cells were counted using Image J. Each cell line is represented by 3 biological replicates. G Cell viability of SUM159PT and Hs578T cell lines treated with PNPLA8 siRNA or scrambled siRNA as control. Each cell line is represented by 3 biological replicates. *p < 0.05, **p < 0.001, ***p < 0.0001