Boletín de Segovia: Número 30 - 1918 marzo 11

Copia digital. Madrid : Ministerio de Cultura. Subdirección General de Coordinación Bibliotecaria, 200

Additional file 5: Figure S3. of The sialyl-glycolipid stage-specific embryonic antigen 4 marks a subpopulation of chemotherapy-resistant breast cancer cells with mesenchymal features

Correlation among CD24-, CD44-, and SSEA4-expressing subpopulations. To address the correlation between CD24, CD44, and SSEA4 expression, we performed costaining of these markers on residual tumor nodules after AC chemotherapy and untreated tumors of three independent models: HBCx-6 (a), HBCx-10 (b), and HBCx-14 (c). Regulation of the three markers did not correlate among the treatment cycles. (PNG 1664 kb

Additional file 9: Table S3. of The sialyl-glycolipid stage-specific embryonic antigen 4 marks a subpopulation of chemotherapy-resistant breast cancer cells with mesenchymal features

Results of microarray analysis. Results of gene and miRNA expression profiling, Gene Ontology analysis, and miRNA target prediction. (XLSX 118 kb

Additional file 1: of The sialyl-glycolipid stage-specific embryonic antigen 4 marks a subpopulation of chemotherapy-resistant breast cancer cells with mesenchymal features

Supplemental experimental procedures. Detailed description of materials and methods. (DOCX 62 kb

Additional file 12: Figure S8. of The sialyl-glycolipid stage-specific embryonic antigen 4 marks a subpopulation of chemotherapy-resistant breast cancer cells with mesenchymal features

SSEA4-positive breast cancer cells show decreased expression of miRNAs inhibiting EMT inducers. Expression ratios of the 12 miRNAs targeting the key mesenchymal regulator and indicator genes ZEB1, ZEB2, fibronectin 1, Snail1, Snail2, and Twist. Each bar represents the log2 expression ratio of the SSEA4-positive fraction relative to the SSEA4-negative fraction for the respective tumor model. (TIFF 201 kb

Additional file 3: Figure S1. of The sialyl-glycolipid stage-specific embryonic antigen 4 marks a subpopulation of chemotherapy-resistant breast cancer cells with mesenchymal features

Representative gating strategy for flow cytometry–based marker analysis of dissociated xenograft tumor tissue. Tumor tissue was dissociated to obtain a single-cell suspension while preserving cell surface epitopes. The sample was stained for mouse-specific markers to exclude cells of murine origin from the analysis as well as for the screening candidates and analyzed by multiparametric flow cytometry. Doublets were excluded by forward scatter (FSC) area/FSC height gating (a); debris was excluded by FSC/side scatter gating (b); dead cells were excluded by gating off propidium iodide–positive events (c); and mouse cells were excluded by gating on α-mouse-fluorescein isothiocyanate–negative events (d). When we screened two samples in parallel, we found that one of the samples was labeled using an ultraviolet dye, allowing for subsequent separation of the events of each sample by gating on the VioBlue channel fluorescence intensity (e–h). (PNG 736 kb

Additional file 13: Figure S9. of The sialyl-glycolipid stage-specific embryonic antigen 4 marks a subpopulation of chemotherapy-resistant breast cancer cells with mesenchymal features

Expression of SSEA4 in RCC and healthy kidney tissue. Primary RCC and healthy kidney tissues from the same patient were dissociated and analyzed by multiparametric flow cytometry. Doublets were excluded by FSC-A/FSC-H gating (a); debris was excluded by FSC/SSC gating (b); dead cells were excluded by gating off PI+ events (c); and lineage-positive cells were excluded by gating on α-Lin-FITC–negative events (d). In each patient, healthy and tumor tissues were analyzed in parallel in one labeling reaction. Therefore, one of the samples was labeled using a UV dye, allowing for subsequent separation of the events of each sample by gating on the VioBlue channel fluorescence intensity (e–h). In all of the analyzed patients (n = 3), the expression of SSEA4 was strongly increased in the tumor tissue as compared with the respective healthy tissue, with almost all tumor cells expressing SSEA4 in two of the patients (f–h). *α-Lin-FITC = CD45-FITC, CD31-FITC, CD235a (glycophorin A)-FITC. (PNG 717 kb