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. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Nature. 2019 Oct 30;575(7782):380–384. doi: 10.1038/s41586-019-1715-0

Figure 4. Multi-tracer PET imaging of KL lung tumors with 18FBnTP and 18F-FDG.

Figure 4.

a, PET/CT images of a KL mouse with two tumors, T1 and T2. Top panel – 18FBnTP/CT overlay, bottom panel – 18F-FDG/CT overlay. H – heart. b, Western blot of T1 and T2 isolated from mouse imaged in a and probed with indicated antibodies. c, Western blot of tumors isolated from KL or KP mice and probed with indicated antibodies. d-i, Experiments performed on A549 (ADC) and RH2 (SCC) cell lines. d, Western blot probed with indicated antibodies. Loading control (β-actin) is shown for each blot; intensity of bands relative to actin is indicated. e, TMRE measurement (n = 3 biological replicates). f, Basal oxygen consumption rate (OCR) (n = technical 15 replicates). g, Extracellular acidification rate (ECAR) (n = technical 15 replicates). h, Coomassie staining of mitochondria isolated from A549 and RH2 cells separated on blue native gel. SC – supercomplex, I – complex I, V – complex V, III – complex III. Densitometry of the CI band corresponding to CV and CIII is indicated. i, Complex I activity in A549 and RH2 cells (n = technical 25 replicates). j, Schematic of the treatment regiment for KL mice. k, Percent of lung area occupied by tumors in KL mice treated with Vehicle or 15 mg/kg IACS-010759. N = 9 mice (Vehicle), n = 13 mice (IACS). l, Percent of Ki67 positive cells per tumor area quantified for TTF1 positive tumors (TTF1POS) for KL mice treated as indicated in j (left panel; n=861 [Vehicle]; n=1068 [IACS]) and CK5 positive tumors (CK5POS) tumors (right panel; n=31 [Vehicle]; n=26 [IACS]). The data are shown as the mean +/− SEM. m, Transverse view of a KL mouse imaged with 18FBnTP PET/CT (top panel) and 18F-FDG PET/CT (bottom panel). H – heart; tumor is outlined. n, H&E stain of the PET imaged tumor. Tumor histology is indicated. Scale bar = 1.0 mm. o, Representative images of Glut1 (left panel) and CK5/TTF1 (right panel) stained tumor. Scale bar = 25 μm. The data are represented as the mean +/− SD. Statistical significance was calculated using unpaired two-tailed t-test. Experiments in a, b, c, d-h were done twice with similar results. Experiments in i, k, l were done once. Data in m-o are representative of three independent mouse experiments.