Figure 2 from N-Myristoytransferase Inhibition Causes Mitochondrial Iron Overload and Parthanatos in TIM17A-Dependent Aggressive Lung Carcinoma
Year: 2024
Type: other
Abstract: NMT inhibition causes mitochondrial ferrous iron accumulation and increases ROS in (KL/K)MUT but not (KL/K)WT lung carcinoma cells. A, TfR1 and DAPI staining on (KL/K)MUT H460 cells treated with 0.5 μmol/L DDD85646 (NMTi) or vehicle for 72 hours. Images were inverted for clarity. Right column, amplification of the area in the square in the middle column. Bar, 15 μm in left and middle columns, 5 μm in right column. (*), endocytic recycling compartment. Arrowheads, intracellular TfR1 clusters. B, Cytoplasmic ferrous iron measured using FerroOrange in H460 cells (KL/K)MUT treated with 0.5 μmol/L DDD85646 (NMTi) or vehicle for the indicated times. Two independent experiments with four technical replicates each were combined. Deferoxamine (DFO, 7 μmol/L) was used as negative control. Fluorescence was normalized to cell number and expressed as arbitrary units (a.u.). Bar, group mean; error bars, SD. **, P = 0.0014; ***, P = 0.0002; ns = not significant (Student t test). C, Mitochondrial ferrous iron (Mito-FerroGreen) measured in (KL/K)MUT and (KL/K)WT lung carcinoma cells treated with 1 μmol/L DDD85646 or vehicle for 24 hours. Signal intensity was quantified in randomly imaged fields containing at least 200 cells per condition. Bar, group mean; error bars, SEM; a.u., arbitrary units. ***, P = 0.0002; ns, not significant (Student t test). Representative images are shown. Bar, 15 μm. D, ROS detected using DCFH-DA in (KL/K)MUT and (KL/K)WT lung carcinoma cells treated with 1 μmol/L DDD85646 for the indicated times. Signal intensity was quantified in randomly imaged fields containing at least 720 cells per condition. Bars, mean; error bars, SEM; a.u., arbitrary units. *, P = 0.0202; ****; P < 0.0001, **; P = 0.0017; ns, not significant (Student t test). Representative images from 72 hours of treatment are shown. Bar, 30 μm.
Language: English
Subfield: Pulmonary and Respiratory Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4400979391