Journal: Translational Oncology

Article Title: Extracellular vesicles extracted from bone marrow mesenchymal stem cells carrying MicroRNA-342-3p inhibit the INHBA/IL13Rα2 axis to suppress the growth and metastasis of breast cancer

doi: 10.1016/j.tranon.2021.101333

Figure Lengend Snippet: The effect of BMSC-EVs carrying miR-342-3p on the proliferation, migration, invasion, and apoptosis of MCF-7 cells. A, The expression of miR-342-3p in the microarray GSE35412. B, The enrichment of miR-342-3p in various tissues determined by EVmiRNA. C, The expression of miR-342-3p in breast cancer cells MCF-7, SKBR3, T47D, MDA-MB-231, and human normal breast epithelial cell line MCF10A determined by RT-qPCR, ** p < 0.01 in comparison with MCF10A cells. D, The entering of PKH67-labeled EVs carrying Cy3-labeled miR-342-3p into MCF-7 cells observed by a confocal microscope. E, The expression of miR-342-3p in MCF-7 cells incubated with BMSC-EVs determined by RT-qPCR. F, MCF-7 cell proliferation determined by CCK-8 assay. G, MCF-7 cell apoptosis detected by flow cytometry. H, Scratch test of MCF-7 cell migration. I, MCF-7 cell invasion detected by Transwell assay. J, The expression of E-cadherin, N-cadherin and Vimentin in MCF-7 cells determined by Western blot analysis. In panel E-J, ** p < 0.01 in comparison with MCF-7 cells incubated with EVs-inhibitor-NC, ## p < 0.01 in comparison with MCF-7 cells incubated with EVs-mimic NC. Cell experiments were conducted three times independently.

Article Snippet: Breast cancer cell lines including MCF-7, SKBR3, T47D and MDA-MB-231, and human normal breast epithelial cell line of MCF10A were purchased from ATCC (Manassas, VA).

Techniques: Migration, Expressing, Microarray, Quantitative RT-PCR, Comparison, Labeling, Microscopy, Incubation, CCK-8 Assay, Flow Cytometry, Transwell Assay, Western Blot

Journal: Translational Oncology

Article Title: Extracellular vesicles extracted from bone marrow mesenchymal stem cells carrying MicroRNA-342-3p inhibit the INHBA/IL13Rα2 axis to suppress the growth and metastasis of breast cancer

doi: 10.1016/j.tranon.2021.101333

Figure Lengend Snippet: The effect of BMSC-EVs-miR-342-3p on the expression of INHBA and IL13Rα2 in MCF-7 cells. A, Venn map of miR-342-3p downstream genes predicted by the mirDIP, DIANA TOOLS, RNA22, TargetScan, and miRWalk databases. B, The top 20 genes interacting of MRFAP1 and INHBA predicted by GeneMANIA. C, The expression data of MRFAP1 in breast cancer obtained from the TCGA database analyzed by GEPIA. D, The expression data of INHBA in breast cancer obtained from the TCGA database analyzed by GEPIA, * p < 0.05. E, The expression of INHBA in breast cancer cells MCF-7, SKBR3, T47D, MDA-MB-231, and human normal breast epithelial cell line MCF10A measured by RT-qPCR (** p < 0.01 in comparison with MCF10A cells). F, The relationship between miR-342-3p and INHBA predicted by TargetScan database and verified by dual luciferase reporter assay in MCF-7 cells, ** p < 0.01 in comparison with mimic-NC. G, The relationship between INHBA and IL13Rα2 detected by MEM analysis. H, Binding between INHBA and IL13Rα2 in MCF7 assessed by RIP. ** p < 0.01 in comparison with MCF7 cells treated with sh-NC. I, IL13Rα2 protein pulled down by INHBA in MCF-7 cells detected by RNA pull-down assay. ** p < 0.01. J, The stability of IL13Rα2 protein in MCF7 cells detected by Western blot analysis. * p < 0.01 in comparison with Control MCF7 cells. K, The expression of INHBA and IL13Rα2 in MCF7 cells determined by Western blot analysis, ** p < 0.01 in comparison with MCF7 cells transfected with mimic-NC. L, The expression of INHBA and IL13Rα2 in MCF7 cells determined by Western blot analysis, ** p < 0.01 in comparison with MCF7 cells treated with EVs-mimic-NC. Cell experiments were conducted three times independently.

Article Snippet: Breast cancer cell lines including MCF-7, SKBR3, T47D and MDA-MB-231, and human normal breast epithelial cell line of MCF10A were purchased from ATCC (Manassas, VA).

Techniques: Expressing, Quantitative RT-PCR, Comparison, Luciferase, Reporter Assay, Binding Assay, Pull Down Assay, Western Blot, Control, Transfection

Journal: Scientific reports

Article Title: Emerin deficiency drives MCF7 cells to an invasive phenotype.

doi: 10.1038/s41598-024-70752-5

Figure Lengend Snippet: Fig. 9. Reduced emerin expression at the nuclear periphery correlates with breast cancer invasiveness in patients. (A) Representative tissue microarray staining of emerin in 159 patients using emerin polyclonal antibodies (Proteintech, cat# 10351-1-AP) or secondary alone (Vector Lab, cat#: MP-7451). Nuclei are blue, emerin is brown, and arrows denote emerin staining in certain images for reference. As severity of cases increases, there is a visible reduction in emerin expression at the nuclear envelope and more deformed nuclei are present. (B) Quantification of emerin staining on IHC-stained patient samples using 0–3, with 0 having no staining at the nuclear periphery and 3 having complete, dark rim staining. N = 159 total samples, *P < 0.05 compared to normal tissue, one-way ANOVA and Dunnett’s test. Error bars represent standard deviation. (C) Representative tissue microarray staining of emerin in 183 patients using emerin monoclonal antibodies (Leica, NCL-Emerin) or secondary alone (Vector Lab, cat#: MP-7452) using the same samples used in A. Nuclei are blue and emerin is brown. As aggressiveness of cases increases, there is a visible reduction in emerin expression and more deformed nuclei are present. (D) Quantification of emerin staining using the 0 to 3 grading system. N = 183 total samples #P < 0.02 compared to all non-cancerous tissue, *P < 0.0062 compared to both normal and benign tissue, one-way ANOVA and Dunnett’s test. Error bars represent standard deviation.

Article Snippet: Reduced emerin expression at the nuclear periphery correlates with breast cancer invasiveness in patients. (A) Representative tissue microarray staining of emerin in 159 patients using emerin polyclonal antibodies (Proteintech, cat# 10351-1-AP) or secondary alone (Vector Lab, cat#: MP-7451).

Techniques: Expressing, Microarray, Staining, Plasmid Preparation, Standard Deviation, Bioprocessing

Journal: PLoS ONE

Article Title: Functional Genomic Analyses of Two Morphologically Distinct Classes of Drosophila Sensory Neurons: Post-Mitotic Roles of Transcription Factors in Dendritic Patterning

doi: 10.1371/journal.pone.0072434

Figure Lengend Snippet: To access differences in gene expression, RNA from enriched C-I and C-IV neuronal populations were compared with that of whole larvae in microarray experiments. Live confocal images of neuronal populations labeled by ( A ) GAL4 221 (C-I) and ( B ) GAL4 ppk1.9 (C- IV) visualized by the trans-membrane fusion construct mCD8::GFP. Neurons have been pseudo-colored to distinguish individual subtypes and dendritic territories. ( C ) GAL4 221 strongly labels C-I da neurons along with weakly labelling of C-IV neurons in the background (dotted red trace). ( D ) GAL80 driven by a ppk promoter was combined in the background of GAL4 221 (ppk-GAL80; GAL4 221 ) that results in highly class I specific GAL4 expression. ( E ) A representative whole larval image of GAL4 ppk1.9 driving the expression of UAS-mCD8::GFP (gut is auto-fluorescent). ( F-J ) Strategy of class-specific neuronal isolation. Larvae expressing mCD8::GFP under the control of either ppk-GAL80; GAL4 221 or GAL4 ppk1.9 ( F ) were dissociated ( G ) filtered and incubated with superparamagnetic beads coated with anti-mCD8 antibody ( H ). The C-I/C-IV neurons bound to the magnetic beads were purified using a strong magnet ( I ), washed several time and used to perform microarray gene expression profiling ( J ). An identical region from the C-I and C-IV microarray are represented to show their dramatic qualitative differences ( J ). The microarray replicates were highly correlated, as represented in the correlation map ( K ). Principle component analysis revealed the three microarray samples from C-I, C-IV and whole larval lysate cluster into three distinct and well-defined clusters ( L ).

Article Snippet: The filtrate is then incubated with superparamagnetic beads (Dynabeads MyOne Streptavidin T1, Invitrogen) coupled with biotinylated mouse anti-CD8a antibody (eBioscience) for 60 minutes.

Techniques: Gene Expression, Microarray, Labeling, Membrane, Construct, Expressing, Isolation, Control, Incubation, Magnetic Beads, Purification

Journal: Cell reports

Article Title: RNA-binding protein FXR1 drives cMYC translation by recruiting eIF4F complex to the translation start site

doi: 10.1016/j.celrep.2021.109934

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: p21 Waf1/Cip1 , Cell Signaling Technology , Cat# 2947, RRID: AB_823586.

Techniques: Control, Recombinant, Modification, Transfection, Protease Inhibitor, CCK-8 Assay, Bicinchoninic Acid Protein Assay, cDNA Synthesis, SYBR Green Assay, Caspase-Glo Assay, Immunoprecipitation, Plasmid Preparation, Staining, In Situ, Microarray, Western Blot, Expressing, Labeling, Negative Control, Mutagenesis, Software

Journal: Cancer Research

Article Title: Slit2 Inhibits Breast Cancer Metastasis by Activating M1-Like Phagocytic and Antifibrotic Macrophages

doi: 10.1158/0008-5472.can-20-3909

Figure Lengend Snippet: Figure 5. Slit2 reduces the expression of IL6, thereby inhibiting TAMs activity. A, Total RNA of CD11bþ cells sorted from rSlit2-N PBS-treated MMTV-PyMT tumorswas subjected to gene expression analysis using NanoString technology. The heatmap shows differentially expressed top genes. B, Total RNA was isolated from MDA-MB-231 cells overexpressing Slit2 (231-Slit2) or vector control (231-vec) and analyzed for gene expression using microarray technology. The heatmap shows differentially expressed top genes. C, 231-Slit2 or 231-Vec cells CM was subjected to cytokine array analysis. The representative image shows differentially expressed molecules. D, The graph shows levels of IL6 in CM derived from 231-Sli2 or Vec CM detected by ELISA. E, Expression of Robo1 in MDA-MB-231 cells transduced with lentivirus expressing siRNA specific to Robo1 (si-Robo1) or control (si-ctrl) by Western blot. F, b-actin was used as loading control. MDA-MB-231 parental control, si-Robo1, or si-ctrl from E was treated with rSlit2-N or PBS and phosphorylation at p65 of NFkB (p-NFkB) or total NFkB (t-NFkB) was analyzed. G, Graph showing levels of IL6 in si-Robo1 or si-ctrl MDA-MB-231 cells treated with rSlit2-N or PBS detected by ELISA (n ¼ 3 each group). H, BMDMs were pretreated with serum-free CM (CTRL), or 231-Slit2 CM or 231-Vec CM or 231-Vec CM preincubated with IL6 nAb (231-Vec þ IL6nAb) or IL6 nAb. After 2 hours, pH-Rhodo–labeled S. Aureus particles were added to the cells and recombinant IL6 was added to the 231-Slit2 CM cells (231-Slit2-rIL6). The kinetics of S. Aureus phagocytosis was analyzed using a fluorescence plate reader at every 15 minutes up to 4 hours. , P < 0.05; , P < 0.01; , P < 0.001 using Student t test.

Article Snippet: The cells were pretreated with mouse rSlit2-N (100 ng/mL) or IL6 neutralization antibody (BioXCell# BE0046) for 2 hours.

Techniques: Expressing, Activity Assay, Gene Expression, Isolation, Plasmid Preparation, Control, Microarray, Derivative Assay, Enzyme-linked Immunosorbent Assay, Transduction, Western Blot, Phospho-proteomics, Labeling, Recombinant