Journal: Breast Cancer : Targets and Therapy

Article Title: Identification of genes involved in breast cancer and breast cancer stem cells

doi: 10.2147/BCTT.S85202

Figure Lengend Snippet: Gene expression among different cancer types. Notes: Relative gene expression analysis of nine genes that were overexpressed in breast cancer and breast CSCs. The gene expression also was studied in colon and lung cancer, colon CSCs, lung CSCs, embryonic stem cells, and in a reference sample. The experiments were performed in triplicate and a P -value of <0.05 was considered significant. Abbreviations: CSCs, cancer stem cells; ESCs, embryonic stem cells; HP , haptoglobin; SMPD1 , sphingomyelin phosphodiesterase 1, acid lysosomal; SCRIB , scribbled homolog ( Drosophila ); KCMF1 , potassium channel modulatory factor 1; FAM155B , family with sequence similarity 155, member B; PTGER3 , prostaglandin E receptor 3 (subtype EP3); GPR3 , G protein-coupled receptor 3; TMX2 , thioredoxin-related transmembrane protein 2; DDX49 , DEAD (Asp-Glu-Ala-Asp) box polypeptide 49.

Article Snippet: During the exponential phase of proliferation, commercial breast CSCs cells were plated in 24-well plates (E36102-29-24Well; Celprogen) and transfected with gene-specific small interfering RNAs (siRNAs) using Lipofectamine 2000 Reagent (11668-027; Thermo Fisher Scientific), according to the manufacturer’s instructions.

Techniques: Expressing, Sequencing

Journal: Breast Cancer : Targets and Therapy

Article Title: Identification of genes involved in breast cancer and breast cancer stem cells

doi: 10.2147/BCTT.S85202

Figure Lengend Snippet: Breast CSCs pre- and post-siRNA knockdown. Note: Representative images showing breast CSCs pre- and post-siRNA knockdown. Abbreviations: CSCs, cancer stem cells; siRNA, small interfering RNA; TMX2 , thioredoxin-related transmembrane protein 2; FAM155B , family with sequence similarity 155, member B; PTGER3 , prostaglandin E receptor 3 (subtype EP3); GPR3 , G protein-coupled receptor 3; DDX49 , DEAD (Asp-Glu-Ala-Asp) box polypeptide 49.

Article Snippet: During the exponential phase of proliferation, commercial breast CSCs cells were plated in 24-well plates (E36102-29-24Well; Celprogen) and transfected with gene-specific small interfering RNAs (siRNAs) using Lipofectamine 2000 Reagent (11668-027; Thermo Fisher Scientific), according to the manufacturer’s instructions.

Techniques: Small Interfering RNA, Sequencing

Journal: Breast Cancer : Targets and Therapy

Article Title: Identification of genes involved in breast cancer and breast cancer stem cells

doi: 10.2147/BCTT.S85202

Figure Lengend Snippet: Gene expression of stemness transcription factors in breast CSCs. Notes: Relative gene expression of transcription factors in breast CSCs following knockdown. The ∆∆Ct method was used to perform the analysis. Each bar represents the average of the Ct values. The assays were performed in triplicate and a P -value of <0.05 was considered to be significant. The assays are presented in a log2 scale. Thus, positive values indicate overexpression while negative values indicate underexpression. Abbreviations: CSCs, cancer stem cells; TMX2 , thioredoxin-related transmembrane protein 2; FAM155B , family with sequence similarity 155, member B; PTGER3 , prostaglandin E receptor 3 (subtype EP3); GPR3 , G protein-coupled receptor 3; DDX49 , DEAD (Asp-Glu-Ala-Asp) box polypeptide 49; NANOG , Homeobox protein NANOG; OCT3/4 , Octamer-binding transcription factor ¾; SOX2 , sex determining region Y-box 2; CD34 , Hematopoietic progenitor cell antigen CD34.

Article Snippet: During the exponential phase of proliferation, commercial breast CSCs cells were plated in 24-well plates (E36102-29-24Well; Celprogen) and transfected with gene-specific small interfering RNAs (siRNAs) using Lipofectamine 2000 Reagent (11668-027; Thermo Fisher Scientific), according to the manufacturer’s instructions.

Techniques: Expressing, Over Expression, Sequencing, Binding Assay

Journal: STAR Protocols

Article Title: Bioprinting of heterogeneous and multilayered cell-hydrogel constructs using continuous multi-material printing and aerosol-based crosslinking

doi: 10.1016/j.xpro.2022.101303

Figure Lengend Snippet:

Article Snippet: Green fluorescent protein (GFP) MDA-MB-231 , ANGIO-PROTEOMIE , CatcAP-0034GFP.

Techniques: Recombinant, Modification, Saline, Clinical Proteomics, Software, Construct, Aerosol

Journal: NPJ Breast Cancer

Article Title: NDRG4 promoter hypermethylation is a mechanistic biomarker associated with metastatic progression in breast cancer patients

doi: 10.1038/s41523-019-0106-x

Figure Lengend Snippet: NDRG4 depletion in breast tumor cells increases lymph node adhesion and cell migration toward VN. a NDRG4 expression meta-analysis in human breast cancer cell lines using GOBO application . Breast cancer subtypes can be identified by different colors: red = Basal A, gray = Basal B and blue = Luminal cells. b Analysis of NDRG4 mRNA expression levels in MCF-7 cells transfected with NDRG4-shRNAs or scramble control (shSCR). Expression levels are relative to wild type cells and normalized to hydroxymethylbilane synthase (HMBS) gene. Error bars represent SEM of biological replicates ( n = 5). *** p < 0.001, ns = not significant, by one-way ANOVA. c Western blot analysis of NDRG4 expression in cytoplasmic extracts of MCF-7 shNDRG4s or shSCRs cells, using antibodies against NDRG4 and Actin. MCF-7 cells express all the three isoforms of NDRG4: 37 kDa (NDRG4-B), 39 kDa (NDRG4-B var ) and 41 kDa isoform (NDRG4-H). For simplicity, results obtained for two independent shNDRG4 clones were grouped and presented as the shNDRG4 group. ( d , left) Representative images of adherent red fluorescent MCF-7 cells on frozen rat lymph node sections. d , e Quantification of adherent MCF7 ( d ) and T47D ( e ) breast tumor cells in lymph nodes sections (right, n = 4 independent experiments for MCF-7 and 3 for T47D, ** p < 0.01 by two-tailed t test). f Stationary adhesion assays showed that NDRG4 knockdown promotes ‘adhesive switch’ between FN and VN. Error bars represent SEM of biological replicates ( n = 4). Cells do not adhere to albumin control (BSA, data not shown). * p < 0.01, ** p < 0.01, ns = not significant by two-tailed t tests. g , h Haptotactic cell migration toward VN was analyzed by transwell migration assay in MCF-7 ( g ) and T47D ( h ) cell lines. Error bars represent SEM of biological replicates ( n = 4). *** p < 0.001 by two-tailed t tests

Article Snippet: For stable transfection of NDRG4 shRNA into MCF-7 and T47D breast tumor cells, the pGFP-V-RS vector containing shRNA inserts which targeted the 3′-unttranslated region of NDRG4 (TRCN0000134583 and TRCN0000137216) was purchased (Origene).

Techniques: Migration, Expressing, Transfection, Western Blot, Clone Assay, Two Tailed Test, Transwell Migration Assay

Journal: NPJ Breast Cancer

Article Title: NDRG4 promoter hypermethylation is a mechanistic biomarker associated with metastatic progression in breast cancer patients

doi: 10.1038/s41523-019-0106-x

Figure Lengend Snippet: NDRG4 knockdown promotes clustering of β1-integrin at the leading edge of T47D cells. Representative confocal images of β1 integrin subunit (MAB1965, green) at the ventral cell surface of VN-adherent T47D shNDRG4 or shSCR cells

Article Snippet: For stable transfection of NDRG4 shRNA into MCF-7 and T47D breast tumor cells, the pGFP-V-RS vector containing shRNA inserts which targeted the 3′-unttranslated region of NDRG4 (TRCN0000134583 and TRCN0000137216) was purchased (Origene).

Techniques: