Supplementary Figures 1 - 13, Tables 1 - 8 from SHON Is a Novel Estrogen-Regulated Oncogene in Mammary Carcinoma That Predicts Patient Response to Endocrine Therapy

PDF file - 1391K, Supplemental Methods Fig. S1. Characterization of a polyclonal anti-SHON antibody raised in rabbits. Fig. S2. Specificity of the rabbit SHON polyclonal antibody. Fig. S3. Forced expression of SHON transforms normal human breast epithelial cells in vitro. Fig. S4. Immunocytochemistry with affinity purified SHON antibody. Fig. S5: Microphotographs of SHON expression in normal and breast cancer tissues. Fig. S6 and S7:: Kaplan-Meier survival curves. Fig. S8. SHON is an estrogen inducible gene. Fig. S9. The effects of endogenous SHON depletion significantly on growth in 3D Matrigel. Fig. S10. Depletion of SHON decreased BCL-2 and NF-kB transcription and protein in MCF-7 cells. Fig. S11. Depletion of endogenous SHON by a second SHON siRNA significantly reduced cell proliferation, anchorage independent growth and migration/invasion in MCF-7 cells. Fig. S12. Oncogenic properties of SHON in T47D mammary carcinoma cells. Fig. S13. BCL2 and NF-κB mediate SHON oncogenicity in T47D cells. Table S1. Primer sequences used to detect gene specific transcripts by RT-PCR. Table S2: The sequences of the primers used for real-time PCR. Table S3: Expression of genes in MCF7-SHON relative to MCF7-Vec cells. Table S4. The sequence of oligonucleotides used to construct SHON siRNA in pSilencer 2.1-U6 hygro vector. Table S5. Clinicopathological characteristics of the whole cohort (n=1650). Table S6: Antigens, primary antibodies, clone, source, optimal dilution and scoring system used for each immunohistochemical marker. Table S7: Association between SHON expression and other clinicopathologic variables. Table S8: Multivariate analysis using Cox regression analysis confirms that SHON protein expression is independent prognostic factor.