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Figure 3. Induction of metastasis of KM-Luc/GFP cells or FM3A-Luc cells to the proper axillary lymph node (PALN) after inoculation of the cells into the subiliac lymph node (SiLN). KM-Luc/GFP cells (A, C, E); FM3A-Luc cells (B, D, F). A: Progression of KM-Luc/GFP cell metastasis from the SiLN to the PALN, as detected from measurements of luciferase activity; B: progression of FM3A-Luc cell metastasis from the SiLN to the PALN, as detected from measurements of luciferase activity; C: quantification of luciferase activity in the SiLN and PALN after inoculation of KM-Luc/GFP cells into the SiLN (SiLN, n = 5 at day 4 and n = 3 at day 7; PALN, n = 4 at day 4 and n = 4 at day 7). ****P < 0.0001, unpaired t-test. The bars show mean ± SEM values; D: quantification of luciferase activity in the SiLN and PALN after inoculation of FM3A-Luc cells into the SiLN (SiLN, n = 14 at day 6, n = 10 at day 13, n = 7 at day 20 and n = 3 at day 27; PALN, n = 14 at day 6, n = 10 at day 13, n = 7 at day 20 and n = 3 at day 27). ***P = 0.0001, ****P < 0.0001, one-way ANOVA and Tukey’s test. The bars show mean ± SEM values; E: representative images of the PALN at day 6, obtained using contrast-enhanced high-frequency ultrasound (CE-HFUS; n = 5; Video 3), in a mouse inoculated with KM-Luc/GFP cells. The CE-HFUS image reveals microbubbles flowing in the vessel in the PALN; F: representative images of the PALN at day 8, obtained using CE-HFUS (n = 5; Video 4), in a mouse inoculated with FM3A-Luc cells. The CE-HFUS image shows microbubbles flowing in the vessel in the PALN. NS: not significant