Depression and family support in breast cancer patients

MTS, migration and invasion assays in DCIS.COM cells that were previously transduced with scrambled control (Control) or BCL9 KD shRNA. The control cells and BCL9 KD cells were re-transduced with empty vector (EV), BCL9 overexpression (BCL9-OE) and BCL9 KD. BCL9-OE was achieved by transduction using the PCDH-BCL9 (BCL9-OE) acquired from Dr. Carrasco [11]. A Western blot analysis was performed using anti-BCL9, anti-vimentin, anti-E-cadherin antibodies, and anti-β-actin as a loading control. B MTS assay on control cells transduced with EV (control + EV), or BCL9-OE (control + BCL9-OE), BCL9-KD transduced with EV (BCL9 KD + EV), and BCL9-KD transduced with BCL9-OE (BCL9 KD + BCL9-OE). Bar graphs represent mean absorbance at 490 nm normalized to control ± standard error of the mean (SEM) (n = 6). C, D Representative images of the migration and invasion assays. Bar graph represents percent area of cells migrated (left) and invaded (right) under the membrane after 24 h. Invasion and migration were determined by ImageJ analysis of microscopic images per sample, the data are mean values normalized to control ± SEM (n = 3). E TopFlash and FopFlash reporter activity in DCIS.COM transduced as above that were either treated with Wnt3A or control conditioned medium (CM). Data represent mean ± SEM (n = 3, letters indicate statistically significant difference). (PDF 964 kb

Supplementary Figure S4 from NEMO, a Transcriptional Target of Estrogen and Progesterone, Is Linked to Tumor Suppressor PML in Breast Cancer

NEMO KD results in decreased IL-6, p-serine STAT3 and PML in vivo.</p

Supplementary Figure S2. from NEMO, a Transcriptional Target of Estrogen and Progesterone, Is Linked to Tumor Suppressor PML in Breast Cancer

NEMO is required for E+P induced expression of IL-6 and NFkB signaling in ER+PR+ overexpressing DCIS.com breast cancer cells.</p

Supplementary Table S1-S3 from NEMO, a Transcriptional Target of Estrogen and Progesterone, Is Linked to Tumor Suppressor PML in Breast Cancer

S1:Differences in response to E+P were not correlated with any available patient data, including biomarker expression, age, or nuclear grade. S2:ER target genes with a role in NFkB pathway.S3:Relapse-free survival (RFS), distant metastasis-free survival (DMFS) and overall survival (OS) in all breast cancer patients or luminal A or B subtypes based on their expression levels of IKBKG and PML</p

Supplementary Figure S3 from NEMO, a Transcriptional Target of Estrogen and Progesterone, Is Linked to Tumor Suppressor PML in Breast Cancer

NEMO KD results in increased cell survival and proliferation in MCF7 cells in vitro.</p

Supplementary Figure S1. from NEMO, a Transcriptional Target of Estrogen and Progesterone, Is Linked to Tumor Suppressor PML in Breast Cancer

Five ER+/PR+ DCIS patient samples were cultured under non adherent conditions and treated with E+P, E, P, and vehicle control. Only a subset of patient-derived ER+/PR+ DCIS cells responded to hormone treatment in vitro by increasing mammosphere formation efficiency.</p

Supplementary Material and Methods from NEMO, a Transcriptional Target of Estrogen and Progesterone, Is Linked to Tumor Suppressor PML in Breast Cancer

Description of additional experiments and procedures carried out in this study</p

Supplementary Figure S5 from NEMO, a Transcriptional Target of Estrogen and Progesterone, Is Linked to Tumor Suppressor PML in Breast Cancer

Kaplan-Meier (KM) relapse-free and overall survival curves of breast cancer patients based on their expression levels of IKBKG, and PML.</p

Additional file 10: of Expression profiling of in vivo ductal carcinoma in situ progression models identified B cell lymphoma-9 as a molecular driver of breast cancer invasion

Analysis of The Cancer Genome Atlas (TCGA) data showing canonical Wnt pathways genes expressed in BCL9-high versus BCL9-low tumors. TCGA breast cancer samples with BCL9 levels above the range defined by normal samples were labeled upregulated in cancer (414 samples). The significant differentially expressed genes from TCGA analysis of BCL9-high versus BCL9-low tumors were analyzed in Ingenuity Pathway Analysis (IPA) and the canonical pathways with a p value ≤0.05 were obtained. (XLS 124 kb

Additional file 5: Figure S2. of Expression profiling of in vivo ductal carcinoma in situ progression models identified B cell lymphoma-9 as a molecular driver of breast cancer invasion

BCL9 showed increased nuclear expression, while BCL9L expression remained cytoplasmic during ductal carcinoma in situ (DCIS) invasive transition. A Immunofluorescence staining of BCL9 (red; top panel), BCL9L (red; bottom panel), K5/K19 (green), and Hoechst (blue) in a primary sample that represents: adjacent normal glands (left), DCIS lesions (middle), and invasive ductal carcinoma (IDC) (right). BCL9 and BCL9L are conjugated to Alexa-Fluor 594 (red) and K5/K19 were conjugated to Alexa-Fluor 488 (green). Nuclei were counterstained with Hoechst. Scale bars 50 μm, × 40 objective was used. B, C RT-qPCR of BCL9 and BCL9L mRNA in epithelial cell adhesion molecule (EpCAM)-positive epithelial cells sorted from SUM225 (B) and DCIS.COM (C) mouse intraductal xenograft model (MIND) xenografts at 2, 6, and 10 weeks post-intraductal injection. The bar graphs represent fold change normalized to 2 weeks. Data are mean values ± standard error of the mean (n = 3, *p <0.05). D Representative western blot analysis of cell lysates from control and BCL9-KD-SUM225 and DCIS.COM blotted with anti-BCL9, and anti-BCL9L antibodies. β-actin was used as a loading control. The analysis showed no change in BCL9L protein levels in BCL9-KD cells compared to control cells. (PDF 5012 kb