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Suominen et al. J Cancer Metastasis Treat 2019;5:14 I http://dx.doi.org/10.20517/2394-4722.2018.64 Page 5 of 13

A

B C

Figure 2. Body weight and MDA-MB231(SA) tumor burden. Whole body tumor burden was analyzed by fluorescence imaging. A:
Representative images showing established metastatic foci before treatment (day 14) and at sacrifice (day 25); B: body weight during
the study relative to body weight at the day of tumor cell inoculation. Significant differences in body weight at sacrifice relative to
baseline were not observed; C: whole body tumor burden did not differ between the groups as measured by fluorescent imaging. Data
are expressed as mean ± SD, n = 8 animals. Scale bars 10 mm, arrows tumor foci in vivo. NS: non-significant; DOX: doxorubicin; ZOL:
zoledronic acid

Anti-tumor effects of DOX, ZOL, and their sequential combination were studied by (1) measuring whole
body tumor burden by fluorescence imaging; and (2) tumor area in bone by histomorphometry [Figures 2C
and 3D, respectively]. According to fluorescence imaging at sacrifice, none of the treatments affected the