Fig. 5

Id2 expression is induced by bone morphogenetic protein (BMP)7 but not transforming growth factor (TGF)β1. a RT-qPCR analysis of Bmp7 and Tgfb1 expression in brains and lungs from naive BALB/c mice (n = 3 independent samples). ***p < 0.001, *p < 0.05, Student’s t test. b Expression levels of Bmp7 in mice cell types isolated from non-tumour-bearing mice in the Srinivasan RNAseq dataset [11] (GSE75246; n = 4 or 5 samples per cell type) and Kamphuis dataset [12] (GSE74614; n = 12 samples per cell type). Scatter plots represent median. *p < 0.05, ***p < 0.001, Mann Whitney test. c RT-qPCR analysis of Id2 expression in 4T1-Luc cells treated with recombinant TGFβ1 (5 ng/mL) or BMP7 (300 ng/mL) for 2–24 h. Data represent 2 wells/condition, mean ± SEM, and are shown relative to the untreated sample. ***p < 0.001, one-way ANOVA. d 4T1-Luc cells were treated with BMP7 (300 ng/mL) for 24 h and cells were washed and incubated in complete media for a further 1 or 5 days before Id2 expression was assessed by RT-qPCR. Data represent 2 wells/condition, mean ± SEM. **p < 0.01, Student’s t test. e,f ID2 expression levels in MDA-MB-231-Luc cells treated as described in b and c. *p < 0.05, one-way ANOVA and Student’s t test. g RT-qPCR analysis of Id1, Id3, and Id4 expression in 4T1-Luc tumour sublines independently isolated from primary tumours, lungs, and brains, relative to Gapdh. p values generated using Mann-Whitney test between tumour and brain. *p < 0.05, **p < 0.01 (lungs were not included in statistical analysis as n = 2 samples). h RT-qPCR analysis of Id1, Id3, and Id4 expression levels in 4T1-Luc cells treated with TGFβ1 (5 ng/mL) or BMP7 (300 ng/mL) for 2–24 h. Data represent 2 samples per time point, mean ± SEM and are shown relative to untreated samples. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. ns not significant