Journal: OncoTargets and therapy

Article Title: PWP1 Promotes the Malignant Phenotypes of Lung Cancer Cells by Interacting with DVL2 and Merlin

doi: 10.2147/OTT.S263815

Figure Lengend Snippet: PWP1 activates the Wnt signaling pathway by interaction with DVL2. After transfection of PWP1 into SK-MES-1 cell line, wb showed that the phosphorylation of DVL2 and Active β-catenin were upregulated, and the expression levels of target genes of Wnt pathway, C-myc, Cyclin D1 and MMP7 were upregulated ( A ). After knocking down of PWP1 into H1299 cell line, wb showed that the phosphorylation of DVL2 and Active β-catenin were downregulated, and the expression levels of target genes of Wnt pathway, C-myc, Cyclin D1 and MMP7 were downregulated ( B ). GAPDH serves as a loading control. The grey value was measured using Image software. Luciferase gene-reporter assays showed that PWP1 could activate the Wnt pathway, after knocking down PWP1, the Wnt pathway would be inhibited ( C ). Columns: mean numbers, Bar: SD. (*P<0.05). PWP1 and DVL2 co-localized in the cytoplasm ( D , magnification 600×). Co-ip testing confirmed the interaction between PWP1 and DVL2 ( E ). Transfection of PWP1 and knocking down DVL2 would offset the effects of PWP1 on the Wnt pathway ( F ). Results are shown from three independent experiments.

Article Snippet: In addition, primary antibodies against PWP1 (#SC-390188, 1: 500), DVL2 (#SC-166303, 1: 500) were purchased from Santa Cruz Biotechnology, Inc. GAPDH (ZSGB-BIO, China, #TA309157, 1: 1000) was also used.

Techniques: Transfection, Phospho-proteomics, Expressing, Control, Software, Luciferase, Co-Immunoprecipitation Assay

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 2 | DVL1 knockdown reduces β-catenin levels in the cartilage and chondrocytes. Mice with OA were administered adenoviral vectors containing either sh-DVL1 or sh-NC via intra-articular injections. Lentiviral vectors carrying sh-DVL1 or sh-NC were introduced into chondrocytes extracted from OA mice. The mRNA (A) and protein (B) levels of DVL1 in chondrocytes were determined using RT-qPCR and WB analysis, respec- tively. (C) Protein levels of β-catenin in chondrocytes determined using WB analysis. (D) The transcriptional activity of β-catenin in chondrocytes analyzed by TOP/FOPFlash assays. (E) Positive staining of DVL1 in the mouse knee joint determined using immunofluorescence staining. (F) mRNA expression of DVL1 in the joint cartilage determined using RT-qPCR. (G) Positive expression of β-catenin in the joint cartilage determined us- ing IHC assay. For animal experiments, each group contained five mice. For cell experiments, three biological replicates were performed. Differences were compared by the unpaired t-test (C, D) or ANOVA (A, B, F, and G).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Knockdown, Quantitative RT-PCR, Activity Assay, Staining, Immunofluorescence, Expressing

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 3 | DVL1 knockdown ameliorates cartilage injury in mice. (A) Protein levels of COL10A1, MMP13, and SOX9 in mouse cartilage tissues determined using WB analysis. (B) Cartilage morphology in the mouse knee joints determined using Safranin O/fast green staining. (C) Positive TRAP staining in the mouse knee joints. (D) Apoptosis in the extracted chondrocytes determined using TUNEL assay. (E) Protein levels of pro- cleaved-caspase-3 in the extracted chondrocytes determined using WB analysis. (F) Protein levels of COL10A1, MMP13, and SOX9 in the mouse chondrocytes determined using WB analysis. For animal experiments, each group contained five mice. For cell experiments, three biological repli- cates were performed. Differences were compared by the unpaired t-test (D–F) or ANOVA (A–C).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Knockdown, Staining, TUNEL Assay

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 4 | CHIR-99021 restores cartilage injury mitigated by DVL1 silencing. Mice stably administered sh-DVL1 were further treated with the Wnt/β-catenin agonist CHIR-99021 via intra-articular injection. (A) Protein levels of β-catenin in cells determined using WB analysis. (B) Cartilage morphology in the mouse knee joints determined using Safranin O/fast green staining. (C) Protein levels of COL10A1 and COL2A1 in mouse chon- drocytes determined using WB analysis. (D) Positive TRAP staining in the mouse knee joints; in vitro, chondrocytes with stable DVL1 knockdown were treated with 5 μM CHIR-99021 for 24 h. (E) Apoptosis in the extracted chondrocytes determined using TUNEL assay. (F) Protein levels of pro- cleaved-caspase-3 in the extracted chondrocytes determined using WB analysis. (G) Protein levels of expression of MMP13 and SOX9 in the mouse chondrocytes determined using WB analysis. For animal experiments, each group contained five mice. For cell experiments, three biological repli- cates were performed. Differences were compared by the unpaired t-test (A–G).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Stable Transfection, Injection, Staining, In Vitro, Knockdown, TUNEL Assay, Expressing

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 5 | SP2, poorly expressed in OA, represses DVL1 transcription. (A) Transcription factor binding sites near the mouse DVL1 promoter region predicted using the JASPAR Transcription Factors plugin from the UCSC Genome Browser. (B) Intersections of the predicted transcription factors and the significant DEGs obtained from high-throughput sequencing. (C) Positive staining of SP2 in the joint cartilage determined using IHC. (D) SP2 mRNA expression in the joint cartilage determined using RT-qPCR. (E) Binding between SP2 and the DVL1 promoter in chondrocytes determined using ChIP-qPCR assay; the extracted chondrocytes were administered lentiviral vectors carrying OE-NC or OE-SP2. (F) mRNA and (G) protein level of SP2 in mouse chondrocytes determined using RT-qPCR and WB analysis. (H) mRNA expression of DVL1 in mouse chondrocytes analyzed using RT-qPCR. (I) Regulation of SP2 on transcription activity of the DVL1 promoter in chondrocytes determined using the dual lucifer- ase reporter gene assay. For animal experiments, each group contained five mice. For cell experiments, three biological replicates were performed. Differences were compared by the unpaired t-test (C–I).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Binding Assay, Next-Generation Sequencing, Staining, Expressing, Quantitative RT-PCR, ChIP-qPCR, Activity Assay, Reporter Gene Assay

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 6 | DVL1 upregulation aggravates cartilage injury in mice alleviated by SP2. DMM-challenged mice were introduced with adenoviral vectors carrying OE-NC/OE-SP2 alone or with the additional OE-NC/OE-DVL1. (A) mRNA expression of SP2 and DVL1 in the joint cartilage deter- mined using RT-qPCR. (B) Positive staining of SP2 and DVL1 in the joint cartilage determined using IHC. (C) Protein levels of β-catenin in the joint cartilage determined using WB analysis. (D) Cartilage morphology in the mouse knee joints determined using Safranin O/fast green staining. (E) Positive TRAP staining of the mouse knee joints. (F) Protein levels of COL10A1 and MMP13 in the mouse cartilage determined using WB analysis. Each group contained five mice. Differences were compared by ANOVA (A–F).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Expressing, Quantitative RT-PCR, Staining

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 7 | DVL1 overexpression increases chondrocyte loss suppressed by SP2. Chondrocytes were administered lentiviral vectors encapsu- lating OE-NC/OE-SP2 or the additional OE-NC/OE-DVL1. (A) mRNA expression of DVL1 in cells determined using RT-qPCR. (B) Apoptosis in cells determined using TUNEL assay. (C) Protein levels of β-catenin in chondrocytes determined using WB analysis. (D) Transcriptional activity of β-catenin in chondrocytes analyzed by TOP/FOPFlash assays. (E) Protein levels of SOX9 and COL2A1 in the chondrocytes determined using WB analysis. Three biological replicates were performed. Differences were compared by unpaired t-test (A) or ANOVA (B–E).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, TUNEL Assay, Activity Assay

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 8 | Schematic illustration of the mechanism. In cartilage tissues of osteoarthritic mice, decreased expression of SP2 resulted in its attenuated transcriptional repression of DVL1. Aberrantly expressed DVL1 activated the β-catenin signaling, leading to chondrocyte damage and osteoarthritis progression.

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Expressing

Journal: bioRxiv

Article Title: The abnormal C-terminus in DVL1 impacts Robinow Syndrome phenotypes

doi: 10.64898/2026.02.14.705933

Figure Lengend Snippet: A) Wild-type human DVL1 with an N-terminal flag tag, followed by the main signaling domains followed by the C-terminal. The DVL1 507fs*141 (DVL1 1519ΔT ) and truncating construct DVL1 507* (DVL1 1519* ) also have an N-terminal FLAG-tag. The nuclear localization sequence (NLS, IxLT) and nuclear export sequence (NES, M/LxxLxL) are maintained in all the constructs. However, the DVL1 1519ΔT construct has a frameshift (fs, red), leading to a novel C-terminus of 141 aa, followed by a STOP codon (asterisk). B) Schematic of RCAS virus injection and skeletal phenotyping. Embryos were injected in the right frontonasal mass at HH15 (embryonic day 2.5) and phenotyping was performed at multiple stages. C) Contingency analysis followed by Wilson/Brown (fraction of total) test showing a statistically significant difference in the fraction of total embryos with abnormal upper beak. D-D’’) Wholemount skulls stained with alcian blue and alizarin red showed a normal patterning of frontonasal mass derived bones (premaxilla, nasal, prefrontal) in GFP-injected specimens (24/24). E-E’’) In contrast, embryos injected with wild-type DVL1 (n=8/14) had a shorter, deviated beak with missing premaxilla. F-F’’) Variant hDVL1 (n=12/13) displayed hypoplastic premaxillary bones. G-G’’) Normal beak formation in all embryos injected with the 1519* truncated variant.. Key: e- eye, fnm – frontonasal mass, fs – frameshift, ios – intraorbital septum, md – mandibular bone, mx- maxillary bone, n- nasal bone, NES – nuclear export sequence, NLS – nuclear localization sequence, ns – nasal slit, p – palatine bone, pmx – premaxilla, Scale bars in D-D’’) apply to all images.

Article Snippet: The open reading frame encoding human DVL1 was purchased from Origene (#RC217691).

Techniques: FLAG-tag, Construct, Sequencing, Virus, Injection, Staining, Derivative Assay, Variant Assay

Journal: bioRxiv

Article Title: The abnormal C-terminus in DVL1 impacts Robinow Syndrome phenotypes

doi: 10.64898/2026.02.14.705933

Figure Lengend Snippet: ( A ) Control wild-type wing with shaded dpp - Gal4 expression domain (green) and black boxes that correspond to zoomed-in views presented in panels ( A’-D’’’ ). ( B-D ) Representative dpp>DVL1- expressing adult female wings at 29°C. ( A’-D’ ) Zoomed in views of PCP defects within a fixed region above the posterior cross vein for control ( A’ ) and DVL1 -expressing ( B’-D’ ) adult female wings ( A’’-D’’ ). Zoomed in views of anterior cross vein (ACV) in control ( A’’ ) and DVL1 -expressing ( B’’-D’’ ) adult female wings. Arrowhead in C’’ points to reduction of ACV. ( A’’-D’’ ) Zoomed in view of adult wing between the L3 and L4 veins in control ( A’’’ ) and DVL1 -expressing (B’’’-D’’’ ) female wings from 29°C crosses where extra bristles and creases are indicated with arrowheads in C’’’ . Phenotypic frequencies are quantified in ( E ). 25 wings were scored per genotype across n=2 independent experiments. Scale bar = 500 μm ( A-D ), 20 μm ( A’-D’ ), 50 μm ( A’’-D’’ ), 100 μm ( A’’’-D’’’ ). ( F ) Western blot analysis of FLAG tagged wildtype, variant and C-terminal truncated DVL1 protein levels from larval head protein extracts. β-tubulin was used as a loading control. ( G ) On the right, plot of DVL1 protein levels in variant DVL1-expressing tissue relative to control ( dpp >wt DVL1 ) tissue. Each dot on the bars represents one blot. Error bars show mean with SD. Statistics were performed with a one-way ANOVA test.

Article Snippet: The open reading frame encoding human DVL1 was purchased from Origene (#RC217691).

Techniques: Control, Expressing, Western Blot, Variant Assay

Journal: bioRxiv

Article Title: The abnormal C-terminus in DVL1 impacts Robinow Syndrome phenotypes

doi: 10.64898/2026.02.14.705933

Figure Lengend Snippet: ( A-A’ ) 29°C control adult female fly phenotypes. ( B-D’ ) Representative dorsal (A-D) and ventral (A’-D’) view images of female adult phenotypes from hh> Gal4 crosses performed at 29°C crossed to (B-B’) wt DVL1, ( C-C’ ) DVL1 1519 Δ T , and ( D-D’ ) DVL1 1519* . Arrowheads in C - C’ points to malformed wings (black) and the legs (white) in hh>DVL1 1519 Δ T flies. 30 flies were collected and observed from n=4 genetic crosses.

Article Snippet: The open reading frame encoding human DVL1 was purchased from Origene (#RC217691).

Techniques: Control

Journal: bioRxiv

Article Title: The abnormal C-terminus in DVL1 impacts Robinow Syndrome phenotypes

doi: 10.64898/2026.02.14.705933

Figure Lengend Snippet: A-C) Histology of embryos that were used for subsequent immunofluorescence experiments in D-L. D-F) Presence of the viral GAG protein is shown with anti-GAG staining in the frontonasal mass There is more staining on the right side of the frontonasal mass since embryos lie with their right side facing upwards in the egg and this is the most accessible site for injection. G-I) Embryos with staining for SOX9 in the mesenchyme. Some areas of relatively higher expression are seen in the globular processes (arrows). J-L) Proliferation in the mesenchyme was high and not affected by the presence of the DVL1 viruses (compared to areas of GAG staining shown in D-F). M) quantification of proliferation in the right half of the frontonasal mass (see schematic) shows no difference between any of the viruses. N) The levels of human DVL1 expression are about 4-5 fold higher than GFP infected controls. There is no significant difference between the wt,DVL1, 1519ΔT and 1519* viruses. O) There is no change in expression of the gallus DVL1 gene so no feedback loops are activated. P) Neither genes in the WNT pathway nor those involved in skeletogenesis were significantly altered in the 1519ΔT variant relative to human wt DVL1 infected frontonasal mass mesenchyme.

Article Snippet: The open reading frame encoding human DVL1 was purchased from Origene (#RC217691).

Techniques: Immunofluorescence, Staining, Injection, Expressing, Infection, Variant Assay

Journal: bioRxiv

Article Title: The abnormal C-terminus in DVL1 impacts Robinow Syndrome phenotypes

doi: 10.64898/2026.02.14.705933

Figure Lengend Snippet: Embryos were injected at stage 15 and fixed 96h post-injection when they had reached stage 29. Near-adjacent sections were used for A,D,G,J,M; B,E,H,K,N; C,F,I,O. Panel L is a different embryo than the others in the right column. A,B) sections stained with Alcian blue and Picrosirius red show differentiating prenasal cartilage in the midline. C) the DVL1 variant has disrupted the cartilage and replaced it with mesenchymal cells. D-F) Broad spread of the virus as shown by GAG staining. G-I) Anti-SOX9 (stains chondrocyte nuclei (red)) and anti-TWIST1 (stains undifferentiated mesenchymal cell nuclei, green). (G, H) In GFP and wild-type hDVL1 infected embryos, SOX9 is expressed in the prenasal cartilage surrounded by undifferentiated TWIST1-positive mesenchyme. I) In embryos infected with hDVL1 1519ΔT variant, TWIST1 replaced SOX9 expression in the prenasal cartilage (right side). J-L , P ) Embryos with BrdU stainin g showed significantly lower proliferation in the hDVL1 1519ΔT infected area compared to the wtDVL1-infected embryos (P). M-O ,Q ) Nuclear β-catenin is expressed in the prenasal cartilage with no significant differences in expression (Q). R) width between the nasal slits is significantly greater in the variant DVL1 infected frontonasal mass compared to GFP or wtDVL1 infected embryos. Key: fnm – frontonasal mass, ns – nasal slit, pnc – prenasal cartilage. Scale bar = 50µm and applies to all panels.

Article Snippet: The open reading frame encoding human DVL1 was purchased from Origene (#RC217691).

Techniques: Injection, Staining, Variant Assay, Virus, Infection, Expressing

Journal: bioRxiv

Article Title: The abnormal C-terminus in DVL1 impacts Robinow Syndrome phenotypes

doi: 10.64898/2026.02.14.705933

Figure Lengend Snippet: A) Mesenchymal cells harvested from the frontonasal mass of stage 24 (E4.5) embryos and were plated into high-density cultures. B) Other cultures were sectioned and used for microscopic analysis. C,E-H) There is a significant decrease in the proportion of cartilage from the DVL1 1519ΔT variant compared to the wtDVL1 infected cultures as measured in wholemount stained cultures. When the DVL1 1519* truncation was compared to wtDVL1 there was no significant difference in the Alcian blue stained area. The DVL1 1519* construct reduced cartilage compared to GFP controls. D,I-L) The cultures were significantly thinner in the presence of the DVL1 1519ΔT variant compared to w tDVL1 or the GFP controls. The DVL1 1519 * construct slightly reduced the thickness of the cartilage compared to GFP controls. M) The viruses containing human DVL1 constructs were expressed at similar levels in primary mesenchyme as determined by qRT-PCR with human-specific DVL1 primers. Generally the human DVL1 gene expression was elevated 10 to 15-fold by the viral transgenesis. N, O) The 1519ΔT virus significantly reduced expression of TWIST2 , MMP13 and LEF1 . P,Q) Two reporters were used in micromass cultures from frontonasal mass cells, the SuperTOPFlash reporter for canonical WNT signaling and ATF2 for JNK-PCP, non-canonical WNT signaling. The wt DVL1 plasmid significantly activated both reporters. In comparison both 1519ΔT and 1519* viruses did not activate the reporters as much as wt DVL1 . They did retain more activity than the control, parent plasmid. Statistical analysis done with one-way ANOVA followed by Dunnett’s multiple comparison test ( M ) or Tukey’s post-hoc test (C,D,N,O,P,Q). Scale bar in E-H = 2 mm, I-L = 20µm.

Article Snippet: The open reading frame encoding human DVL1 was purchased from Origene (#RC217691).

Techniques: Variant Assay, Infection, Staining, Construct, Quantitative RT-PCR, Gene Expression, Virus, Expressing, Plasmid Preparation, Comparison, Activity Assay, Control

Journal: bioRxiv

Article Title: The abnormal C-terminus in DVL1 impacts Robinow Syndrome phenotypes

doi: 10.64898/2026.02.14.705933

Figure Lengend Snippet: ( A-D ) Z-stack maximum projection of imaginal wing discs showing DNA (DAPI, blue), the expression of the genetically encoded Jnk reporter puc-lacZ (β-gal; white) and UAS transgene expression domain (red) staining in control ( A ) and DVL1 -expressing female wing discs ( B-D ). ( A’-D’ ) Z-stack maximum projection of single channel images showing β-gal (white) staining in control ( A’ ) and DVL1 -expressing female wing discs ( B’-D’ ). n numbers in bottom panels depict the number of wing discs that displayed the phenotype shown in the representative image. Crosses were performed at 29°C and 11-14 female wing discs were imaged and quantified per genotype across n=3 independent experiments. Scale bar = 50 μm. ( E ) Plot of puc-lacZ puncta counted by finding signal intensity maxima in each wing disc imaged. Error bars show mean with SD. Statistics were performed with ANOVA. ( F-I ) Z-stack maximum projection of third instar wing imaginal discs showing Mmp1 (white) and dpp>GFP expression domain (green) in control ( F ) and DVL1 -expressing female wing disc pouches ( H-I ). ( F’-I’ ) Z-stack maximum projection of single channel images showing Mmp1 staining in control ( F’ ) and DVL1 -expressing female wing discs ( H’-I’ ). Scale bar = 50 μm. ( J ) Plot showing the number of Mmp1 puncta counted per wing disc. Crosses were performed at 29°C and 18 female wing discs were analyzed per genotype. Statistics were performed with ANOVA. ( K-N ) Z-stack maximum projections of Arm protein (white) and dpp>GFP expression domain (green) staining in control (dpp>GFP,+) ( K ) and DVL1-expressing female wing discs ( L-N ) with the corresponding single channel Arm (white) staining shown in ( K’-N’ ). Significant decrease in Arm levels relative to control is indicated with white arrow in M’. Scale bar = 50 μm. ( O ) Schematic of an imaginal wing disc pouch with dpp-Gal4 expression domain (green), position of most stabilized Arm protein (white). Boxes 1 and 2 show the regions where ratio of Arm signal intensity was quantified within and outside of the dpp expression domain. ( P ) The Arm signal intensity ratio quantified as described is plotted. Crosses were performed at 29°C and analyses show averages from 12-16 discs per genotype. Error bars show mean with SD. Statistics were performed with ANOVA.

Article Snippet: The open reading frame encoding human DVL1 was purchased from Origene (#RC217691).

Techniques: Expressing, Staining, Control