Journal: Cell death & disease

Article Title: Long non-coding RNA DSCR8 acts as a molecular sponge for miR-485-5p to activate Wnt/β-catenin signal pathway in hepatocellular carcinoma.

doi: 10.1038/s41419-018-0937-7

Figure Lengend Snippet: Fig. 7 DSCR8 activates Wnt/β-catenin pathway by DSCR8/miR-485-5p/FZD7 axis. Rescue experiments revealed that DSCR8 negatively regulated the miR-485-5p expression (a, b), whereas positively regulated both the mRNA (a, b) and protein (c, d) expression of FZD7 by DSCR8/miR-485-5p/ FZD7 axis. c, d Western blot results from rescue experiments showed that DSCR8 positively regulated the accumulation of cytoplasmic β-catenin and the accumulation of nuclear β-catenin, c-Myc expression, and cyclin D1 expression by DSCR8/miR-485-5p/FZD7 axis. n = three repeats with similar results, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: FZD7 Human cDNA ORF Clone (FZD7) and FZD7 siRNA (si-FZD7) were purchased from OriGene (OriGene Technologies,Inc., USA).

Techniques: Expressing, Western Blot

Journal: Cell death & disease

Article Title: Long non-coding RNA DSCR8 acts as a molecular sponge for miR-485-5p to activate Wnt/β-catenin signal pathway in hepatocellular carcinoma.

doi: 10.1038/s41419-018-0937-7

Figure Lengend Snippet: Fig. 9 Schematic of the proposed mechanism of DSCR8 in HCC. DSCR8 acts as a molecular sponge for miR-485-5p to regulate FZD7 expression, subsequently activating Wnt/β-catenin signal pathway to promote HCC progression

Article Snippet: FZD7 Human cDNA ORF Clone (FZD7) and FZD7 siRNA (si-FZD7) were purchased from OriGene (OriGene Technologies,Inc., USA).

Techniques: Expressing

Journal: The FASEB Journal

Article Title: Mapping of Wnt-Frizzled interactions by multiplex CRISPR targeting of receptor gene families

doi: 10.1096/fj.201700144R

Figure Lengend Snippet: Constructs used in this study

Article Snippet: FZD7 , pCMV6-XL5 FZD7 , Origene , SC122259 , .

Techniques: Construct, Luciferase

Journal: Cell death & disease

Article Title: Long non-coding RNA DSCR8 acts as a molecular sponge for miR-485-5p to activate Wnt/β-catenin signal pathway in hepatocellular carcinoma.

doi: 10.1038/s41419-018-0937-7

Figure Lengend Snippet: Fig. 7 DSCR8 activates Wnt/β-catenin pathway by DSCR8/miR-485-5p/FZD7 axis. Rescue experiments revealed that DSCR8 negatively regulated the miR-485-5p expression (a, b), whereas positively regulated both the mRNA (a, b) and protein (c, d) expression of FZD7 by DSCR8/miR-485-5p/ FZD7 axis. c, d Western blot results from rescue experiments showed that DSCR8 positively regulated the accumulation of cytoplasmic β-catenin and the accumulation of nuclear β-catenin, c-Myc expression, and cyclin D1 expression by DSCR8/miR-485-5p/FZD7 axis. n = three repeats with similar results, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: FZD7 Human cDNA ORF Clone (FZD7) and FZD7 siRNA (si-FZD7) were purchased from OriGene (OriGene Technologies,Inc., USA).

Techniques: Expressing, Western Blot

Journal: Cell death & disease

Article Title: Long non-coding RNA DSCR8 acts as a molecular sponge for miR-485-5p to activate Wnt/β-catenin signal pathway in hepatocellular carcinoma.

doi: 10.1038/s41419-018-0937-7

Figure Lengend Snippet: Fig. 9 Schematic of the proposed mechanism of DSCR8 in HCC. DSCR8 acts as a molecular sponge for miR-485-5p to regulate FZD7 expression, subsequently activating Wnt/β-catenin signal pathway to promote HCC progression

Article Snippet: FZD7 Human cDNA ORF Clone (FZD7) and FZD7 siRNA (si-FZD7) were purchased from OriGene (OriGene Technologies,Inc., USA).

Techniques: Expressing

Journal: Cell Death & Disease

Article Title: Long non-coding RNA DSCR8 acts as a molecular sponge for miR-485-5p to activate Wnt/β-catenin signal pathway in hepatocellular carcinoma

doi: 10.1038/s41419-018-0937-7

Figure Lengend Snippet: a Data from bioinformatics tools (microRNA.org, TargetScan, and miRDB) showed that there were putative binding sites between 3′UTR of FZD7-wt and miR-485-5p. FZD7-mut means mutation of binding sites in 3′UTR of FZD7. b Luciferase reporter gene assays revealed that miR-485-5p negatively regulated the luciferase activity of FZD7-wt-3′UTR, rather than of DSCR8-mut-3′UTR. The mRNA ( c , d ) and protein ( e , f ) expression of FZD7 was negatively regulated by miR-485-5p. And FZD7 clone or siRNAs reversed the effects of miR-485-5p mimics or inhibitors on FZD7 expression ( c – f ). e , f Western blot results revealed that the accumulation of cytoplasmic β-catenin and the accumulation of nuclear β-catenin, c-Myc expression, and cyclin D1 expression were negatively regulated by miR-485-5p, while reversed by FZD7 clone and siRNA. n = three repeats with similar results, ** P < 0.01, *** P < 0.001

Article Snippet: FZD7 Human cDNA ORF Clone (FZD7) and FZD7 siRNA (si-FZD7) were purchased from OriGene (OriGene Technologies,Inc., USA).

Techniques: Binding Assay, Mutagenesis, Luciferase, Activity Assay, Expressing, Western Blot

Journal: Cell Death & Disease

Article Title: Long non-coding RNA DSCR8 acts as a molecular sponge for miR-485-5p to activate Wnt/β-catenin signal pathway in hepatocellular carcinoma

doi: 10.1038/s41419-018-0937-7

Figure Lengend Snippet: Rescue experiments revealed that DSCR8 negatively regulated the miR-485-5p expression ( a , b ), whereas positively regulated both the mRNA ( a , b ) and protein ( c , d ) expression of FZD7 by DSCR8/miR-485-5p/FZD7 axis. c , d Western blot results from rescue experiments showed that DSCR8 positively regulated the accumulation of cytoplasmic β-catenin and the accumulation of nuclear β-catenin, c-Myc expression, and cyclin D1 expression by DSCR8/miR-485-5p/FZD7 axis. n = three repeats with similar results, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: FZD7 Human cDNA ORF Clone (FZD7) and FZD7 siRNA (si-FZD7) were purchased from OriGene (OriGene Technologies,Inc., USA).

Techniques: Expressing, Western Blot

Journal: Cell Death & Disease

Article Title: Long non-coding RNA DSCR8 acts as a molecular sponge for miR-485-5p to activate Wnt/β-catenin signal pathway in hepatocellular carcinoma

doi: 10.1038/s41419-018-0937-7

Figure Lengend Snippet: DSCR8 acts as a molecular sponge for miR-485-5p to regulate FZD7 expression, subsequently activating Wnt/β-catenin signal pathway to promote HCC progression

Article Snippet: FZD7 Human cDNA ORF Clone (FZD7) and FZD7 siRNA (si-FZD7) were purchased from OriGene (OriGene Technologies,Inc., USA).

Techniques: Expressing

Journal: bioRxiv

Article Title: Cachd1 is a novel Frizzled- and LRP6-interacting protein required for neurons to acquire left-right asymmetric character

doi: 10.1101/2022.05.16.492129

Figure Lengend Snippet: (A, left) Representative scatter plot of flow cytometry testing binding of FLAG-tagged CACHD1 prey protein to human FZD7-eGFP transiently transfected HEK293E cells (detected by phycoerythrin (PE)-conjugated secondary antibody; A, right) without (blue) or with (red) pre-incubation with anti-Frizzled antibody OMP-18R5; secondary only negative control (grey). n = 3; one-tailed paired t -test (D. F. = 2, t = 9.53, *** p = 0.0054). (B) Dot plot of human (blue diamonds) or zebrafish (blue circles) CACHD1, or negative control CD200R (grey) prey protein binding (∆M PE ) to cells transiently transfected with eGFP fusion protein constructs indicated (transfections verified by antibody labelling in bold). Each dot represents a single experiment; horizontal bars denote the mean and error bars represent 95% confidence intervals. One way Welch test of means (Cachd1 prey v. CD200R prey, not assuming equal variances; F = 132.32, D. F num = 30.00, D. F denom = 34.67, p = 5.09 × 10 −28 ), post hoc pairwise t -tests with non-pooled standard deviations, Benjamini & Hochberg correction for multiple testing; only significant differences between Cachd1 and CD200R prey for individual transfections are presented here for clarity, ** 0.05 > p > 0.01, *** 0.01 > p > 0.005, **** p < 0.005. (C) SPR-based determination of K D for mouse CACHD1 ECD analyte binding to immobilised FZD5 CRD , LRP6 P3E3P4E4 (3-4, left panel), and normalised response curves for different CACHD1 ECD :FZD CRD interactions. RU, response units.

Article Snippet: Mouse Fzd5 cysteine rich domain (UniProt: Q9EQD0, residues A27-T157), human FZD7 CRD (UniProt: O75084, residues Q33-G170), human FZD8 CRD (UniProt: Q9H461, residues A28-T158) were synthesized (Genscript).

Techniques: Flow Cytometry, Binding Assay, Transfection, Incubation, Negative Control, One-tailed Test, Protein Binding, Construct