Extended Data Figure 7. In vivo assessment of the functional importance of GluN2B-NMDAR signaling in brain-metastatic human breast cancer cells.

a-b, Representative immunofluorescent images of FMRP (green), luciferase (red) and DAPI (blue) in brain metastases formed by shCTRL and shGRIN2B MDA231 cells (a), and quantification of mean FMRP fluorescence in Luciferase⁺ tumour cell clusters (b). Two-tailed Student t-test, mean ± s.e.m.; n = 25 for shCTRL and n = 32 shGRIN2B group, all from 3 mice per group.

c, Weight of orthotopic breast tumors formed by MDA231-BrM cells transfected with inducible shCTRL or shGRIN2B inoculated into the 4th mammary fat pads of female mice. DOX food was added to induce the shRNAs concomitantly with MFP injection. Two-tailed Student t-test, mean ± s.e.m.; n = 9 mice for shCTRL group, and 10 mice for shGRIN2B group.

d, Bio-luminescent imaging (BLI) and quantification of lung metastatic lesions formed by MDA231-BrM cells transfected with inducible shCTRL or shGRIN2B inoculated IV. DOX food was added to induce the shRNAs concomitantly with IV injection. Two-tailed Student t-test, mean ± s.e.m.; n = 5 mice per group, two independent experiments.

e, Bio-luminescent imaging (BLI) and quantification of brain metastatic lesions formed by MDA231-BrM cells transfected with inducible shCTRL or shGRIN2B, or additionally with rescue expression of a GRIN2B cDNA. DOX food was supplied concomitantly with ICD injection to induce shRNA expression. Tumor burden was assessed 4 weeks later by BLI. Two independent experiments. Two-tailed Student t-test, mean ± s.e.m.; n = 10 mice for shCTRL group, 9 mice for shGRIN2B group and 9 mice for rescue group.