Fig 1.
ROS scavenger-NAC reversed oridonin inhibited cell proliferation and oridonin induced intracellular ROS production in oesophageal cancer KYSE-150 cells.
(A) Effects of oridonin on the viability of KYSE-150 cells. (B) DCFH-DA assay of the effects of NAC on oridonin induced ROS production in KYSE-150 cells. (C) Statistical analysis of the effects of NAC on oridonin induced ROS production in KYSE-150 cells. (D) Effects of ROS scavenger-NAC on oridonin inhibited viability of KYSE-150 cells, *p<0.05, **p<0.01, ***p<0.001.
Fig 2.
ROS scavenger-NAC reversed oridonin induced oesophageal cancer KYSE-150 cell apoptosis.
(A) Annexin V/PI assay of the effects of NAC on oridonin induced KYSE-150 cell apoptosis. (B) Statistical analysis of the effects of NAC on oridonin induced KYSE-150 cell apoptosis, ***p<0.001.
Fig 3.
ROS scavenger-NAC reversed oridonin induced disruption of mitochondrial membrane potential in oesophageal cancer KYSE-150 cells.
(A) Rhodamine 123 assay of the effects of NAC on oridonin induced disruption of mitochondrial membrane potential in KYSE-150 cells. (B) Statistical analysis of the effects of NAC on oridonin induced disruption of mitochondrial membrane potential in KYSE-150 cells, **p<0.01.
Fig 4.
ROS scavenger-NAC reversed oridonin induced cell morphology damage of oesophageal cancer KYSE-150 cells.
AFM morphology imaging of (A) control, (B) 10 μM oridonin treated, (C) 30 μM oridonin treated, (D) 50 μM oridonin treated, (E) 2.5 mM NAC+50 μM oridonin treated and (F) 2.5mM NAC treated KYSE-150 cells. (A1-F1) Topography images and (A2-F2) their corresponding 3D images of KYSE-150 cells; (A3-F3) Enlarged topography images in (A1-F1) and (A4-F4) their corresponding 3D images of KYSE-150 cells, scale bar: 20 μm.
Fig 5.
ROS scavenger-NAC reversed oridonin induced oesophageal cancer KYSE-150 cell membrane ultrastructural changes.
AFM morphology and membrane ultrastructure imaging of (A) control, (B) 10 μM oridonin treated, (C) 30 μM oridonin treated, (D) 50 μM oridonin treated, (E) 2.5 mM NAC+50 μM oridonin treated and (F) 2.5mM NAC treated KYSE-150 cells. (A1-F1) Topogrphy images of KYSE-150 cells, scale bar: 20 μm; (A2-F2) Enlarged membrane ultrastructure images in (A1-F1) and (A3-F3) their corresponding 3D images of KYSE-150 cells, scale bar: 500 nm; (A4-F4) Height distribution and roughness of cell surface ultrastructure analyzed from (A2-F2).
Fig 6.
Statistical results of NAC reversed oridonin induced oesophageal cancer KYSE-150 cell membrane ultrastructural changes determined by AFM.
(A) Height distribution, (B) root-mean-squared roughness (Rq) and (C) average roughness (Ra) analyzed from 2×2 μm frame ultrastructure images of KYSE-150 cells, n = 30, *p<0.05, ***p<0.001.
Fig 7.
ROS scavenger-NAC reversed oridonin induced changes of Young’s modulus in oesophageal cancer KYSE-150 cells.
Typical Young’s modulus maps obtained from (A) control, (B) 10 μM oridonin treated, (C) 30 μM oridonin treated, (D) 50 μM oridonin treated, (E) 2.5 mM NAC+50 μM oridonin treated and (F) 2.5mM NAC treated KYSE-150 cells.
Fig 8.
ROS scavenger-NAC reversed oridonin induced changes of Young’s modulus in oesophageal cancer KYSE-150 cells.
(A) Histogram distribution of Young’s modulus obtained from KYSE-150 cell. (B) Statistical analysis of the effects of NAC on oridonin induced KYSE-150 cell Young’s modulus changes, n>5000, ***p<0.001.
Fig 9.
Schematic diagram showing the ROS-mediated oridonin induced KYSE-150 cell apoptosis that can be detected by AFM.