Journal: International journal of molecular sciences

Article Title: MiR-21 Is Required for the Epithelial-Mesenchymal Transition in MDA-MB-231 Breast Cancer Cells.

doi: 10.3390/ijms22041557

Figure Lengend Snippet: Figure 5. The silencing of miR-21 reduced the mesenchymal phenotype of MDA-MB-231 cells. (A) Protein levels of epithelial and mesenchymal markers were tested by Western blotting. β-actin was used as a loading control. The densitometry graph represents protein expression levels of band intensities obtained from at least three independent experiments. The relative band intensities of each sample were calculated by normalizing against β-actin. Each data point represents average ± SD. **** p < 0.0001. (n = 3) (B–E). Immunofluorescence assay was performed to show E-cadherin levels and localization, Snail, Zeb-1, and vimentin (green). (F–G) Cancer stem cell markers ALDH1, GATA2 expressions (red), and localizations were assessed by using immunofluorescence in MDA-MB-231 wt and miR-21 KO cells. ToPro3 (blue) was used for nuclei. KO2 immunofluorescence results were shown as a representative to compare to wt (n = 3), scale bar 20 nm.

Article Snippet: The wells were washed with PBS twice and primary antibodies E-cadherin, Zeb1, Snail (20C8) (Invitrogen, Waltham, MA, USA), vimentin and GSK3B (Cell Signaling Technology; Danvers, MA, USA) ALDH1 (B-5) and GATA2 (H6) Santa Cruz Biotechnology; CA, USA), Wnt-11 (GeneTex; CA, USA) added and incubated for one hour.

Techniques: Western Blot, Control, Expressing

Journal: Genes & Development

Article Title: miRNAs cooperate in apoptosis regulation during C. elegans development

doi: 10.1101/gad.288555.116

Figure Lengend Snippet: The 3′ UTR of egl-1 is a target of miR-35 and miR-58 family miRNAs in vivo. ( A ) The 3′ UTR of egl-1 is illustrated in blue, with a reported C. elegans Argonaut ALG-1-binding site from a recent study indicated above . Sequences are given for predicted miR-35 family-binding sites and miR-58 family - binding sites, and conserved bases within each site are highlighted in gray across four Caenorhabditis species. The mutated sequences corresponding to egl-1 mut mir-35 (red) and egl-1 mut mir-58 (purple) are shown (for complete mutated sequences, see the Materials and Methods). ( B ) A schematic representation of the 3′ UTR reporters constructed for this study. ( C ) Analysis of single-copy 3′ UTR reporter expression during embryogenesis. The genetic background and transgene under investigation are indicated at the left of each image sequence, which shows representative embryos from three developmental stages. For each embryo, a DIC image ( left ) and GFP image ( right ) are shown. The following alleles were used: bcSi25 [P mai-2 gfp::h2b::mai-2 3′ UTR ], bcSi26 [P mai-2 gfp::h2b::egl-1 wt 3 ′ UTR], bcSi27 [P mai-2 gfp::h2b::egl-1 mut mir-35 3′ UTR ], bcSi46 [P mai-2 gfp::h2b::egl-1 mut mir-35 3′ UTR ], and bcSi47 [P mai-2 gfp::h2b::egl-1 mut mir-35 mir-58 3′ UTR ]. Asterisks indicate autofluorescence. Transgenic strains were homozygous for unc-119(ed3) and the cb-unc-119(+) selection marker. Bars, 10 µm.

Article Snippet: Second, this P mai-2 fragment was fused to the 1287-bp coding sequence of gfp::h2b as amplified from the plasmid pCM1.35 (a gift from G. Seydoux; Addgene plasmid no. 17248) using the primers 5′-CTAATTCACTCAATTTTCAGAATGAGTAAAGGAGAAGAACTTTT-3′ and 5′-GCTCGCGTTCTTGTACTGCAAATTACTTGCTGGAAGTGTACTTG-3′.

Techniques: In Vivo, Binding Assay, Construct, Expressing, Sequencing, Transgenic Assay, Selection, Marker

Journal: Scientific Reports

Article Title: Evaluation of uterine receptivity after gonadotropin releasing hormone agonist administration as an oocyte maturation trigger: a rodent model

doi: 10.1038/s41598-019-48918-3

Figure Lengend Snippet: Endometrial PR signalling following oocyte maturation trigger. ( A ) Distribution of PR in endometrium during pre-implantation. Scale bars represent 100 μm. ( B ) Ihh , ( C ) Areg , and ( D ) Gata2 mRNA expression during pre-implantation. Error bars represent standard error of the mean. Bars labelled with distinct letters are significantly different from each other ( P < 0.05).

Article Snippet: Next, mRNA levels of the following genes were quantified with TaqMan Gene Expression Assay (Life Technologies): Hsd3b1 (Mm01261921_mH), Cyp19a1 (Mm00484049_m1), Ihh (Mm00439613_m1), Areg (Mm01354339_m1), Gata2 (Mm00492301_m1), Lif (Mm00434762_g1), Muc1 (Mm00449604_m1), Ltf (Mm00434787_m1), and Foxa2 (Mm01976556_s1).

Techniques: Expressing