Supplementary Figure 13: Characterization of 4T1-luc tumor xenografts (parent and GLUT1-knockout lines).

(a) Western blot analysis of GLUT1 expression in excised tumors. As expected, GLUT1 was present in 4T1-luc but not in 4T1-luc-GLUT1^-/- no. 1. (b) Immunohistochemical staining of excised 4T1-luc and 4T-luc-GLUT1^-/- no. 1 tumors after in vivo non-invasive imaging with BiGluc probe. Tumors were stained with an antibody selectively recognizing GLUT1 protein. Scale bar = 100 µm. (c) Total photon flux from tumor-bearing mice injected with 100 µM of D-luciferin in the presence and absence of GLUT1 specific inhibitor (WZB-117). The data indicate that the presence of the inhibitor does not have any effect on the light output from tumors. Data are represented as mean ± SD (n = 4 per group, biologically independent experiment). (d) Average tumor volume before ¹⁸F-FDG PET/CT imaging. No significant difference between groups was observed. Data are represented as mean ± SD (n = 10 per group, biologically independent experiment). P values were calculated using two-tailed t-test, ns – p>0.05. All experiments were performed at least twice.