Journal: PLoS ONE

Article Title: Expression Profiling of Stem Cell-Related Genes in Neoadjuvant-Treated Gastric Cancer: A NOTCH2 , GSK3B and β-catenin Gene Signature Predicts Survival

doi: 10.1371/journal.pone.0044566

Figure Lengend Snippet: Genes analysed by TaqMan® low density array.

Article Snippet: FZD1 , 8321 , Hs00268943_s1 , 83.

Techniques:

Journal: Research

Article Title: Isovalerylspiramycin I Reprograms the Immunosuppressive and Temozolomide-Resistant Microenvironment by Inhibiting the Frizzled-5/Wnt/β-Catenin Pathway in Glioblastoma

doi: 10.34133/research.0828

Figure Lengend Snippet: ISP-I targets FZD5 to inhibit the Wnt/β-catenin signaling pathway. (A) RNA-seq analysis indicated that ISP-I alters the transcriptome of LN-18. Volcano plot depicting the distribution and top 10 differentially expressed genes. (B) KEGG pathway enrichment analysis revealed classification and functional annotation of enriched pathways. The horizontal axis represented the number of genes, and the vertical axis showed the classification entries. (C) KEGG analysis highlighted significant suppression of Wnt/β-catenin signaling among altered pathways. (D) Normalized transcript expression values (nTPM) for CTNNB1 were calculated in GBM cell lines from the Human Protein Atlas project. (E) Changes in β-catenin levels were determined through Western blotting. (F) qRT-PCR analysis of CCND1 and Myc mRNA levels under ISP-I treatment ( n = 3). (G) Protein levels of cyclin D1 and c-Myc were measured by Western blotting. (H) The effect of ISP-I on levels of core proteins within the Wnt/β-catenin signaling pathway was measured by Western blotting in LN-18 cells. (I) The result of the molecular docking of ISP-I with FZD5 using Autodock Vina was visualized and simulated by PyMOL software. Interaction mode simulation of ISP-I and FZD5 (FZD5: green, ISP-I: yellow, interface residues: pink). (J) ISP-I increased the FZD5 thermal stability as assessed by temperature-dependent CETSA ( n = 3). (K) SPR assays were performed to quantify the binding affinity of ISP-I to FZD5. (L) Confocal microscopy images of FZD5 (red) and fluorescein-hydrazide-labeled ISP-I (green). Pearson’s coefficient was calculated from scatterplots (ImageJ). Scale bars = 10 μm. Data: mean ± SD; significance was calculated with one-way ANOVA test. ** P < 0.01; *** P < 0.001.

Article Snippet: An anti-FZD5 antibody (68204-H001, Sino Bio, Beijing, China) was used as a positive control.

Techniques: RNA Sequencing, Functional Assay, Expressing, Western Blot, Quantitative RT-PCR, Software, Binding Assay, Confocal Microscopy, Labeling

Journal: Research

Article Title: Isovalerylspiramycin I Reprograms the Immunosuppressive and Temozolomide-Resistant Microenvironment by Inhibiting the Frizzled-5/Wnt/β-Catenin Pathway in Glioblastoma

doi: 10.34133/research.0828

Figure Lengend Snippet: ISP-I targets FZD5, thereby inhibiting the Wnt/β-catenin signaling pathway. This inhibition leads to the suppression of PD-L1 and MGMT expression, augmenting glioblastoma cell susceptibility to TMZ and reprogramming the immunosuppressive microenvironment.

Article Snippet: An anti-FZD5 antibody (68204-H001, Sino Bio, Beijing, China) was used as a positive control.

Techniques: Inhibition, 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: Primers used for indel-nested PCR 1 for validation of mutations generated by sgRNAs

Article Snippet: FZD1 , pCMV-XL4 FZD1 , Origene , SC117910 , .

Techniques: Generated

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: FZD1 , pCMV-XL4 FZD1 , Origene , SC117910 , .

Techniques: Construct, Luciferase

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: TaqMan qPCR primers and probes

Article Snippet: FZD1 , pCMV-XL4 FZD1 , Origene , SC117910 , .

Techniques:

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: Multiple targeting of FZD homologous genes with single sgRNA

Article Snippet: FZD1 , pCMV-XL4 FZD1 , Origene , SC117910 , .

Techniques: Mutagenesis

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: Multiple targeting of up to 5 FZD homologous genes with single sgRNA for FZD1,2,4,5,7,8 sgRNA

Article Snippet: FZD1 , pCMV-XL4 FZD1 , Origene , SC117910 , .

Techniques:

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: Mapping of canonical Wnt-FZD interactions. A , C ) FZD1,2,7 sgRNA-I#1 cells were transfected with indicated Wnt and FZD plasmids together with TCF4/Wnt–firefly luciferase and actin– Renilla reporters. Results of 3–4 independent experiments are shown as means ± sem . B ) Several FZDs rescue induction of active β-catenin levels upon Wnt3 expression in FZD1,2,7 sgRNA-I#1 cells. Cells were transfected with Wnt3 and indicated FZDs plasmids for 48 h. Representative experiment out of 3 independent experiments is shown.

Article Snippet: FZD1 , pCMV-XL4 FZD1 , Origene , SC117910 , .

Techniques: Transfection, Luciferase, Expressing