Figure 2. Gluc_tag multiplex assay in vivo.

(a) U87-FM cells expressing different Gluc_tag were implanted subcutaneously in nude mice (n = 3) and tumor growth was monitored overtime. Shown are representative Fluc bioluminescence images of the U87-FM-Gluc_tag-CFP tumors expressing the Gluc_Flag reporter. (b), Quantitation of the different U87-FM tumors expressing different Gluc_tag reporters; Fluc activity (black), calliper measurement (green), total Gluc_tag activity in the blood (red), and bound Gluc_tag activity after immunobinding (blue). (c–d) U87-FM cells expressing the different Gluc_tag reporters were mixed in equal amounts and injected subcutaneously in the same mouse (n = 5). Tumor growth was monitored using Fluc imaging and total Gluc blood assay as in (a–b). Individual cells subpopulation within the same tumor were monitored using the multiplex Gluc_tag immunobinding assay. (e) Immunostaining for the ten different tags expression in the mixed U87-FM-Gluc_tag-CFP tumor. Black arrows indicate positive staining for cells expressing the corresponding Gluc_tag. Size bar = 200 μm. N.d. = not detectable. (f–g) Mixed population of U87-FM cells expressing all ten different tags were implanted intracranially in the brain of nude mice (n = 5). Tumor growth was monitored with Fluc imaging and Gluc total blood assay, and different tumor cell subpopulations were monitored using the multiplex Gluc_tag assay as in (c–d). Fluc bioluminescence imaging of a representative mouse is shown in (f). Data shown in (b,d, and g) as average relative light units (RLU) ± standard deviation. Heatscales in (a, c and f) indicate relative photon levels (photons/sec) as measured by the CCD camera.