Supplementary Fig 1 from Heme Sequestration Effectively Suppresses the Development and Progression of Both Lung Adenocarcinoma and Squamous Cell Carcinoma
Supplementary Figure 1. The levels of proteins and enzyme subunits involved in OXPHOS and glutamine metabolism increase with lung tumor progression. A, Representative H&E and IHC images of lung tissue sections from KLLuc mice showing levels of the OXPHOS protein HCCS. B, Representative H&E and IHC images of lung tissue sections from KLLuc mice showing levels of the glutamine transporter SLC1A5. C, Representative H&E and IHC images of lung tissue sections from KLLuc mice showing levels of GLS. Shown are montage images of lung tissue sections stained with H&E (row 1), montage images stained with DAPI (row 2) or antibodies against the indicated protein (row 3), and 10X images of lung tissue sections stained with DAPI (row 4) or antibodies against the indicated protein (row 5). The light green lines in DAPI montage images outline the tumors in the lung. The blue rectangles in H&E and white rectangles in DAPI montage images denote the regions shown in 10X. Scale bar, montage, 1mm; 4X, 200 µm; 10X, 100µm. D, Graph showing quantified levels of HCCS correlating with A. E, Graph showing quantified levels of SLC1A5 correlating with B. F, Graph showing quantified levels of GLS correlating with C. Protein levels were quantified, and data are plotted as mean {plus minus} SEM. G, Representative 40X IHC images of lung tissue sections showing the co-localization of ATP5F1A, COX7B, and CYCS with the mitochondrial marker TOMM20. Scale bar, 10 µm. H, OCR of NSCLC H226 cells cultured in normal medium, or heme depleted medium with 0.5 mM succinyl acetone (HD+SA). Heme (5 µM, 10 µM, and 20 µM, respectively) was added back to examine the effects of heme on OCRs. For statistical analysis, the levels in cells with heme added back were compared with the levels in cells without heme added back with a Welch two-sample t test. **, P<0.005.