Figure 4
From: Elucidation of in vitro cellular steps induced by antitumor treatment with plasma-activated medium
PAM treatment induced loss of plasma cell integrity and apoptosis. (A) Plasma membrane integrity observed thanks to propidium iodide (red) penetration. Scale bar: 100 µm. (B) Quantification by videomicroscopy of propidium iodide fluorescence in spheroid over the time after exposition to non-activated medium (black circles), PAM (red squares) or gas alone treatment (blue triangles). (C) Quantification by flow cytometry of apoptosis (Annexin-V positive/propidium iodide PI negative cells) through early externalization of phosphatidylserine in cells isolated from spheroids exposed for 1 h, 4 h or 6 h to PAM. (D) Caspases 3/7 activity observed by wide-field fluorescence microscopy after PAM or gas alone treatment or in control condition. (E) Propidium iodide penetration and caspases 3/7 activity observed by wide-field fluorescence microscopy 24 h after exposure to PAM in cohesive spheroid’s core. Crown of detached cells was removed by gentle pipetting. Scale bar: 100 µm. **p < 0.005, ns: non-significant.
