Fig. 2: LAPTM4B suppresses ferroptosis in NSCLC.

A KEGG analysis of the top 30 pathways regulated by LAPTM4B. Left Panel: Analysis from wild-type (WT) and LAPTM4B knockout (KO) A549 cells. Right Panel: Analysis from WT and LAPTM4B KO H1299 cells. B Flow cytometry analysis of lipid peroxidation using a fluorescence-based reporter BODIPY® 581/591 C11. Left panel: Representative experiment showing oxidized ROS levels in A549 and A549 KO cells. Right panel: Quantification of oxidized ROS levels from three independent experiments, presented as mean ± SEM. Data normalized to “A549”. p(A549, A549 KO) = 0.0000001. C Flow cytometry analysis of lipid peroxidation using a fluorescence-based reporter BODIPY® 581/591 C11. Left panel: Representative experiment showing oxidized ROS levels in H1299 and H1299 KO cells. Right panel: Quantification of oxidized ROS levels from three independent experiments, presented as mean ± SEM. Data normalized to “H1299”. p(H1299, H1299 KO) = 0.0263. D Measurement of malondialdehyde (MDA) levels in WT and KO A549 cells, as well as in WT and KO H1299 cells. Quantification of MDA levels from four independent experiments, presented as mean ± SEM. p(A549, A549 KO) = 0.0336. p(H1299, H1299 KO) = 0.027. E Measurement of the glutathione disulfide (GSSG)/glutathione (GSH) ratio and the amount of GSSG normalized to total protein in WT and KO A549 cells. Quantification of GSSG/GSH ratio and GSSG/Protein levels from three independent experiments, presented as mean ± SEM. For GSSG/GSH, p(A549, A549 KO) = 0.0029. For GSSG/Pro, p(A549, A549 KO) = 2.68E−06. F Transmission electron microscopy analysis of mitochondria ultrastructure in WT and KO A549 cells. Upper panel: Representative images showing mitochondria morphology. Lower panel: Quantification of mitochondria size, elongated mitochondria, and damaged mitochondria from four independent experiments, presented as mean ± SEM. Data normalized to “A549”. For mitochondria size, p(A549, A549 KO) = 0.0021. For elongated mitochondria, p(A549, A549 KO) = 0.0131. For damaged mitochondria, p(A549, A549 KO) = 0.0078. Scale bar: 1 µm. The region in the dashed red box is amplified in the lower panel. G Transmission electron microscopy analysis of mitochondria ultrastructure in WT and KO H1299 cells. Upper panel: Representative images showing mitochondria morphology. Lower panel: Quantification of mitochondria size, elongated mitochondria, and damaged mitochondria from four independent experiments, presented as mean ± SEM. Data normalized to “H1299”. For mitochondria size, p(H1299, H1299 KO) = 0.0001. For elongated mitochondria, p(H1299, H1299 KO) = 0.0247. For damaged mitochondria, p(H1299, H1299 KO) = 0.0133. Scale bar: 1 µm. The region in the dashed red box is amplified in the lower panel.