Fig. 6

Intracellular reactive oxygen species (ROS) production regulates PgLPS-induced autophagy. HaCaT cells were either treated with 10 μM PgLPS alone or 10 μM PgLPS plus 100 μM N-acetylcystein (NAC) for 24 h. Cells treated with medium alone were control (Ctr). a Representative fluorescence images are shown to demonstrate an intracellular ROS production (red) using CellROX reagents. Nuclei were stained with Hoechst 33324 (blue). Bar = 50 μm. b Western blotting assay was performed with antibodies against phosphorylated AMPK (p-AMPK), AMPK, Bcl-2, beclin-1 (BECN-1), or LC3. β-actin (ACTB) was used as a loading control. Band densities were presened as antibodies’ expression fold-increases (normalized to ACTB) and compared with the results for untreated cells (Ctr). Quantification results are shown below the corresponding blots. All experiments were performed in quadruplicate