Fig. 5

Targeting AST factors suppress cancer cell dissemination and metastasis. A Representative images of MDA-MB-231-mock versus 4AST cell morphology. Scale bar, 50 μm. B Schematic overview of mammary fat pad xenografts of 4AST factor depleted LM2 cells. C Representative images of primary tumors dissected after injection of mock or 4AST factor depleted LM2 cells. D Measurement of primary tumor growth in LM2-mock versus 4AST factor-depleted tumors. n = 3; ns, not significant. E Measurement of EpCAM⁺/CD45⁻ CTC counts normalized by total blood cells from duplicated slides harvested from control and 4AST factor-depleted LM2 baring mice. F Bioluminescence ex vivo images of lung metastases after LM2 fat pad injection. G-H Measurement of lung metastases quantified by the number of metastatic lung nodules derived from mock versus AST factor-depleted LM2 cells. I Kaplan–Meier survival plot showing the survival rates of mice injected with mock or 4AST factor-depleted LM2 cells, n = 10. J Images of HEK293A-AST^TetR cells co-treated with pomalidomide (100 μM) or lenalidomide (50 μM) after doxycycline-mediated AST induction for 3 days. Scale bar, 50 μm. K Blockade of IKZF1 accumulation in doxycycline-induced HEK293A-AST^TetR cells co-treated with pomalidomide or lenalidomide. L Schematic of the orthotopic breast tumor model subjected to intraperitoneal injection of pomalidomide (10 mg/kg) or lenalidomide (10 mg/kg) once every 3 days. M Kaplan–Meier survival plot showing the survival rates of LM2-injected mice treated with lenalidomide (10 mg/kg). Control, n = 10; lenalidomide; n = 10. N–O Images of primary tumors (N) and lungs (O) dissected from mice treated with either vehicle or lenalidomide. P Measurement of lung metastases by vehicle versus lenalidomide administration quantified by number of metastatic lung nodules. Q Kaplan–Meier survival plot showing the survival rates of LM2-injected mice treated with pomalidomide (10 mg/kg). Control, n = 10; pomalidomide; n = 15. R-S Images of primary tumors (R) and lungs (S) dissected from mice treated with either vehicle or pomalidomide. T Representative images of B16F10-mock versus 4AST cell morphology. Scale bar, 50 μm. U Representative images of primary tumors resulting from B16F10 cell injection into mouse footpad. Above, B16F10-control primary tumor; Below, B16F10-Ikzf1^−/−-derived primary tumor; Scale bar, 0.5 cm. V A bar plot showing the numbers of circulating tumor cells (CTCs) in B16F10-control and -IKZF1^−/−-injected mouse blood, respectively. Error bars are means ± SD of n = 3 independent replicates. *p < 0.05. W A bar plot showing the numbers of CTCs in vehicle and lenalidomide-treated mouse blood, respectively. Error bars are means ± SD of n = 3 independent replicates. *p < 0.05. X–Y Images of primary tumors (X) and lungs (Y) dissected from mice treated with either vehicle or lenalidomide