Fig. 3. The disruption of the ALDOA–actin filament interaction stimulates NOX1-mediated elevation of ROS and Ca²⁺ levels in cancer cells.

a The UM0112176-induced partial dissociation of ALDOA from β/γ actin filaments in vitro. The values are given as a mean and SD, *p < 0.05. b The impact of various combinations of UM0112176, ALDOA and cofilin on the depolymerization of β/γ actin. c UM0112176-induced ALDOA dissociation causes F-actin bundling and depolymerization in KLN205 cells. d Docking result of UM0112176 to ALDOA using Autodock. Inhibitor UM0112176, and residues C72, K293, and C338 are shown in stick style, while the ALDOA fold is shown in cartoon style. All oxygen atoms are colored in red, carbon in green, sulfur atoms in yellow, and nitrogen atoms are colored in blue. Charges on the electrostatic potential surface of ALDOA are represented in blue for regions of positive charge and red for regions of negative charge. Figure elaborated with PyMol (The PyMOL Molecular Graphics System, Version 2.0 Schrödinger, LLC.) e, f Apocynin diminishes UM0112176-induced ROS levels in KLN205 and hNSCLC cells, respectively. g NOX1 silencing diminishes ROS levels in KLN205 cells treated with UM0112176. h NOX1-associated fluorescent signal (green) before and after its silencing in KLN205 cells. Nuclei were counterstained with DAPI (blue). Bar = 20 µm. The graph on right shows a decrease in NOX1-associated fluorescence after silencing. i Apocynin partially protects rise in calcium levels after UM0112716 treatment. j The effect of apocynin on caspase 3 activation. The image shows the activated caspase 3 (green) in KLN205 cells. Bar = 20 µm. k Apocynin prevents disruption of the actin cytoskeleton in cells treated with UM0112176. Bar = 20 µm