Supplementary Figure 1: ROR1 promotes proliferation and mobility of breast cancer cells.
From: A ROR1–HER3–lncRNA signalling axis modulates the Hippo–YAP pathway to regulate bone metastasis
(a) Oncomine box plot showing ROR1 copy numbers in human ERPR-/HER2- triple negative breast cancer (TNBC) and non-TNBC subtypes (n = 73 and 495 tumors respectively, one-way ANOVA). (b) Oncomine box plots showing ROR1 expression levels in a series of human TNBC (n = 211, 178, 46, 26, 22 and 39 tumors, one-way ANOVA) and non-TNBC subtypes (n = 1,340, 320, 250, 89, 44 and 129 tumors, one-way ANOVA). (c) Oncomine box plots showing ROR1 expression levels in human Non-small Cell Lung Carcinoma (n = 33, 63, 10, 26, and 132 tumors, one-way ANOVA) and Small cell lung carcinoma (n = 9 and 6 tumors, one-way ANOVA) or squamous cell lung carcinoma cohorts (n = 154, 75, 10, 26, and 21 tumors, one-way ANOVA). (d and e) Immunoblotting (IB) of indicated proteins in parental/ROR1 KO MDA-MB-231 (d) and BoM-1833 (e) single cell clones. (f and g) Cell proliferation assay of individual (f) and pooled (g) clones of ROR1 KO MDA-MB-231 cells. (h) IB detection of indicated proteins in ROR1 KO cells expressing WT ROR1 or K506A mutant. (i) Cell invasion assay of ROR1 KO MDA-MB-231 cells with expression of indicated plasmid (left, Scale bars, 200 μm; right, quantification). (j and k) Cell migration (j) and invasion (k) assay of pooled clones of ROR1 KO MDA-MB-231 cells with overexpression of indicated plasmid (Scale bars, 200 μm). (l) TRAP staining showing the osteoclast differentiation in the presence of M-CSF only, M-CSF + RANKL, or combined M-CSF + RANKL and conditioned media (CM) from ROR1 KO BoM-1833 cells or CM from ROR1 KO BoM-1833 cells KO supplemented with PBS or recombinant CTGF (50 ng ml−1) (Scale bars, 200 μm). For a-c, the boxes show the median ± 1 quartile, with whiskers extending to the most extreme data point within 1.5 interquartile ranges from the box boundaries. For f, g and i-k, mean ± s.e.m. were derived from n = 3 independent experiments (n.s., P > 0.05, ∗P < 0.05 and ∗∗P < 0.01, ∗∗∗P < 0.001, two-tailed paired Student’s t-test). Unprocessed original scans of all blots with size marker are shown in Supplementary Fig. 9 Statistics source data for i-l are in Supplementary Table 8.
