Data Supplement from ID1 Promotes Breast Cancer Metastasis by S100A9 Regulation

Supplementary Figure 1. Expression of Id1 and Id2 was determined by qRT-PCR in MCF7 cells expressing a vector control, or Id1, or Id2. Supplemenatary Figure 2. Id1 expression in human primary breast cancer and metastatic samples, and MCF7 cells expressing a vector control or Id1 was determined by qRT-PCR (A). Immunoblot of Id1 expression in MCF7 cells expressing a vector control or Id1 (B). Supplementary Figure 3. Top gene candidates that are suppressed by Id1 but not by Id2 are listed. The fold change shows the expression fold decrease in MCF7 cells stably expressing Id1 versus control. Heatmap shows the expression of top ten genes across samples with blue shades indicating lower expression and red shades indicating higher expression of a gene in a sample. Supplementary Figure 4. Id1 suppresses S100A9 expression. (A) Quantitative RT-PCR of S100A9 expression in MCF7 cells transiently transfected with a vector control, or Id1, or Id2. (B) Immunoblots of S100A9 expression in MCF7 cells transiently transfected with a vector control, or Id1, or Id2. Supplementary Figure 5. S100A9 mediates the Id1 functions in migration and invasion. (A) S100A9 expression in MCF7 cells transiently transfected with a vector control, or Id1, or Id1 and S100A9. (B-C) MCF7 cells transiently transfected with a vector control, or Id1, or Id1 and S100A9 were subjected to migration assay (B) and invasion assay (C). Id1 expression in MCF7 cells increased cell migration and invasion. Expression of S100A9 cDNA reversed migratory and invasive phenotypes induced by Id1 expression (B and C). Supplementary Figure 6. Cell growth of MCF7 cells expressing a vector control, or Id1, or Id2, or Id1 and S100A9, or Id1 plus S100A9 and RhoC. Expression of Id1, or Id2, or S100A9 or RhoC in MCF7 cells did not affect cell growth.