Fig. 5: Knockdown of CALCB or RAMP1 inhibits proliferation of Ewing sarcoma (EwS) cells in vivo.

a Left panel: Analysis of tumor growth of A673 EwS cells with/without dox-induced knockdown of CALCB in NSG mice (n = 33). Event was defined as average diameter of 15 mm. Event-free survival time of mice was analyzed by the Kaplan–Meier method and a log-rank test. Right panel: Knockdown of CALCB in the tumors of dox-treated mice was verified by qRT-PCR. Given are mean normalized gene expression levels and SEM; unpaired two-tailed Student’s t test. b Left panel: Analysis of tumor growth of A673 EwS cells with/without dox-induced knockdown of RAMP1 in NSG mice (n = 10). Event was defined as average diameter of 15 mm. Event-free survival time of mice was analyzed by the Kaplan–Meier method and a log-rank test. Right panel: Knockdown of RAMP1 in the tumors of dox-treated mice was verified by qRT-PCR. Given are mean normalized gene expression levels and SEM; unpaired two-tailed Student’s t test. c Histological analysis of the number of mitoses in tumor tissue of EwS xenografts with/without dox-induced knockdown of CALCB or RAMP1. Given is the mean number of mitoses and SEM per high-power filed (HPF) of 22 representative A673/TR/shCALCB xenografts shown in a and 10 A673/TR/shRAMP1 xenografts shown in b. Unpaired two-tailed, Student’s t test. d Histological analysis of necrosis in tumor tissue of EwS xenografts with/without dox-induced knockdown of CALCB or RAMP1. Given is the average percentage of necrotic area and SEM of 22 representative A673/TR/shCALCB xenografts shown in a and 10 A673/TR/shRAMP1 xenografts shown in b. Unpaired two-tailed, Student’s t test. n.s. P > 0.05; **P ≤ 0.01; ***P ≤ 0.001