Journal: PLoS ONE

Article Title: Definition of Critical Periods for Hedgehog Pathway Antagonist-Induced Holoprosencephaly, Cleft Lip, and Cleft Palate

doi: 10.1371/journal.pone.0120517

Figure Lengend Snippet: Single doses of vismodegib were administered at discrete time points indicated by tick marks on the x-axis, including: GD7.0, 7.25, 7.5, 7.75, 8.0, 8.25, 8.5, 8.625, 8.75, 8.875, 9.0, 9.25, 9.5, 9.75, and 10.0. Cyclopamine was administered by subcutaneous infusion from GD8.25 to ~9.375. Representative examples of distinct face and palate phenotypes are shown, including apparently normal (Normal), HPE, CL/P, and CPO. Note that lateral lip clefts resulting from acute vismodegib exposure typically extended into the primary palate (D’), while those resulting from cyclopamine exposure extended into both the primary and secondary palate (F’). The penetrance of HPE, CL/P, and CPO phenotypes resulting from stage-specific vismodegib exposure is shown in the graph. 5–7 litters were examined for each exposure permutation.

Article Snippet: Vismodegib (Toronto Research Chemicals) was suspended as 3mg/ml in 0.5% methyl cellulose (Sigma) with 0.2% Tween (Sigma).

Techniques:

Journal: PLoS ONE

Article Title: Definition of Critical Periods for Hedgehog Pathway Antagonist-Induced Holoprosencephaly, Cleft Lip, and Cleft Palate

doi: 10.1371/journal.pone.0120517

Figure Lengend Snippet: Superior views of dissected brains are shown for a vehicle-exposed normal animal (A) and for representative examples of animals with vismodegib-induced HPE (B), cyclopamine-induced CL/P (C), and vismodegib-induced CPO (D). Severe hypoplasia of the cerebral cortices (cc) and olfactory bulb (ofb) absence is apparent in the animal with HPE. In animals with CL/P and CPO, the cerebral cortices appear to be of approximately normal size but the olfactory bulbs are hypoplastic. Serial coronal sections of comparably classified animals are shown in E-P. Notable HPE-associated features include a single central nasal passage with nasal septum (ns) cartilage absence (black arrow), olfactory bulb agenesis (J), and a single telencephalic vesicle (N). Grossly normal division of the olfactory bulbs (K, L) and cerebral cortices (O, P), and apparent forebrain septal (s) region hyperplasia (white arrows) is observed in animals with CL/P and CPO. (t) Tongue, (e) eye, (sp) secondary palate.

Article Snippet: Vismodegib (Toronto Research Chemicals) was suspended as 3mg/ml in 0.5% methyl cellulose (Sigma) with 0.2% Tween (Sigma).

Techniques:

Journal: PLoS ONE

Article Title: Definition of Critical Periods for Hedgehog Pathway Antagonist-Induced Holoprosencephaly, Cleft Lip, and Cleft Palate

doi: 10.1371/journal.pone.0120517

Figure Lengend Snippet: In vehicle-exposed embryos at GD14.5 (A) the secondary palatal shelves have approximated and made contact at the midline. In affected cyclopamine-exposed embryos with cleft lip (B), palatal shelves are widely spaced and deficient in width. In vismodegib-exposed embryos (C), secondary palatal shelves have also elevated but are deficient in both length and width. Length (D) and width (E) measurements (arbitrary units), as depicted by the dashed calipers, were made on light microscopy images . Shelf width was determined at 1/3 shelf length from the most rostral aspect. *** p<0.001, **** p<0.0001

Article Snippet: Vismodegib (Toronto Research Chemicals) was suspended as 3mg/ml in 0.5% methyl cellulose (Sigma) with 0.2% Tween (Sigma).

Techniques: Light Microscopy

Journal: PLoS ONE

Article Title: Definition of Critical Periods for Hedgehog Pathway Antagonist-Induced Holoprosencephaly, Cleft Lip, and Cleft Palate

doi: 10.1371/journal.pone.0120517

Figure Lengend Snippet: Along with a vehicle-exposed control (A), representative examples of phenotypic outcomes are shown with numbers indicating the gestational stage of acute vismodegib administration. Later exposure was associated with forelimb ectrodactyly, as exhibited bilaterally in fetuses exposed from 9.25 to 9.75 (arrows point to absent fifth digits on the right limb). Kinked tail phenotypes were caused by exposure between GD9.5 and 10.0 (arrowheads). Edema is also apparent in fetuses exposed at GD9.75 and 10.0. For each treatment group the number of litters and fetuses examined, mean litter size and crown-rump length, and the incidence of edema, forelimb ectrodactyly, and kinked tail defects are presented in .

Article Snippet: Vismodegib (Toronto Research Chemicals) was suspended as 3mg/ml in 0.5% methyl cellulose (Sigma) with 0.2% Tween (Sigma).

Techniques: Control

Journal: Nature Communications

Article Title: Identification of recurrent FHL2-GLI2 oncogenic fusion in sclerosing stromal tumors of the ovary

doi: 10.1038/s41467-019-13806-x

Figure Lengend Snippet: a Cell titer blue proliferation assay of immortalized mesenchymal stem cells (MSCs), HEK-293, medulloblastoma (DAOY) and human basal cell carcinoma (BCC) cells stably expressing empty vector (control), wild-type FHL2 (FHL2), wild-type GLI2 (GLI2), truncated GLI2 (tGLI2) or FHL2-GLI2 treated with 250 nM Vismodegib or vehicle control (DMSO). b Representative images of colony formation assay of MSC, HEK-293, DAOY, and BCC cells stably expressing control, FHL2, GLI2, tGLI2, or FHL2-GLI2 treated with Vismodegib 500 nM or vehicle control (DMSO). Scale bars, 5 mm. Quantification of the number of colonies/well compared to control (bottom). c Wound healing assay of MSC, HEK-293, DAOY, and BCC cells stably expressing control, FHL2, GLI2, tGLI2, or FHL2-GLI2 treated with 250 nM Vismodegib or vehicle control (DMSO). The migratory effect/wound area was assessed at 0 and 24 h and quantified compared to DMSO (bottom). Vismo, Vismodegib. Scale bars, 500 μm. In a – c , data are representative of at least three independent experiments. Error bars, s.d. of mean; n.s., not significant; * P < 0.05, ** P < 0.01, *** P < 0.001; two-tailed unpaired t -test.

Article Snippet: The smoothened inhibitors Cyclopamine (Selleckchem, S1146), Vismodegib (Cellagen tech, C4044–5) and the GLI inhibitor GANT61 (Tocris, 3191) were resuspended in DMSO and used in in vitro proliferation, colony formation and scratch wound healing assays at 5–10 μM, 250–500 nM, and 5–10 μM, respectively.

Techniques: Proliferation Assay, Stable Transfection, Expressing, Plasmid Preparation, Control, Colony Assay, Wound Healing Assay, Two Tailed Test

Journal: Molecular Cancer

Article Title: Hedgehog signaling drives glial cell plasticity and oncogenic reprogramming in gastroenteropancreatic neuroendocrine neoplasms

doi: 10.1186/s12943-026-02611-y

Figure Lengend Snippet: Patient-derived PanNET tumoroids express HH signaling proteins and respond to HH pathway activation and inhibition. A Combined fluorescence and phase-contrast images of 21-day-old human PanNET tumoroids (PanNET3) stained for chromogranin A (CHGA) and HH proteins (PTCH1, SHH). B Four patient-derived PanNET tumoroid lines (PanNET1–4) were exposed to recombinant human SHH N-terminal peptide (100 ng/mL) or the SMO inhibitor vismodegib (VISMO, 20 µM) for 72 h and changes in mRNA expression were analyzed by RT-qPCR. mRNA changes were normalized to HPRT1 expression and DMSO vehicle control. ( n = 4 unique patient lines). * = p < 0.05, ** = p < 0.01, **** = p < 0.0001 by Two-way ANOVA with Sidak post-test. C EdU labeling showing proliferation of dissociated PanNET3 cells following 5-day exposure to: the HH agonists SHH-N (100 ng/mL) and SAG (10 nM); inhibitors of the canonical HH signaling pathway vismodegib (20 µM) and sonidegib (10 nM); or inhibitors of the GLI1/2 effectors GANT61 (10 µM) and itraconazole (ITZ, 1 µM). D Quantitation of the percentage of EdU-positive PanNET tumor cells following 5-day treatment. ( n = 3 replicates from one patient tumoroid line). * = p < 0.05, by One-way ANOVA with Tukey post-test. E SHH or SAG were co-administered with the respective pharmacologic inhibitors and EdU uptake was evaluated after 7 days. F Immunofluorescent images of CHGA, SHH, and PTCH1 expression in a second PanNET tumoroid line (PanNET5). G , H EdU labeling was evaluated in PanNET5 tumoroids following 7-day treatment with SHH-N (200 ng/mL) and SAG (20 nM); inhibitors of the canonical HH signaling pathway vismodegib (20 µM) and sonidegib (10 nM); or inhibitors of the GLI1/2 effectors GANT61 (10 µM) and itraconazole (ITZ, 5 µM). I Crystal violet staining of human BON-1 PanNET cells after 48 h treatment. J BrdU incorporation in BON-1 cells after 48 h treatment with HH agonists SHH-N and SAG, ( K ) SMO inhibitors vismodegib and sonidegib, and ( L ) inhibitors of GLI1/2 signaling. M Immunofluorescent images of CHGA, SHH, and PTCH1 expression in tumoroids derived from a metastatic ileal NET (IL-NET-met1). N , O EdU labeling was assayed in the IL-NET-met1 tumoroid line following 7-day treatment with the same drug concentrations used for PanNET5. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test

Article Snippet: Drug compounds used in the studies include: human recombinant SHH N-terminal peptide (R&D Systems, Cat# 1845-GMP), SAG (Tocris, Cat# 4366), vismodegib (Tocris, Cat# 7710), sonidegib (Tocris, Cat# 7826), GANT61 (Tocris, Cat# 3191), and itraconazole (Tocris, Cat# 5981).

Techniques: Derivative Assay, Activation Assay, Inhibition, Fluorescence, Staining, Recombinant, Expressing, Quantitative RT-PCR, Control, Labeling, Quantitation Assay, BrdU Incorporation Assay

Journal: Molecular Cancer

Article Title: Hedgehog signaling drives glial cell plasticity and oncogenic reprogramming in gastroenteropancreatic neuroendocrine neoplasms

doi: 10.1186/s12943-026-02611-y

Figure Lengend Snippet: HH signaling regulates the growth of GFAP ΔMen1 and Sox10 ΔMen1 pancreatic NET tumoroids. A Phase contrast and ( B ) fluorescence images of PanNET tumoroids from Sox10-Cre; Men1 FL/FL ; LSL-tdTomato mice. Tumoroids were imaged after 72 h exposure to: the HH agonists SHH-N (100 ng/mL) and SAG (10 nM); inhibitors of the canonical HH signaling pathway vismodegib (20 µM) and sonidegib (10 nM); or inhibitors of the GLI1/2 effectors GANT61 (10 µM) and itraconazole (ITZ, 1 µM). C TdTomato fluorescence intensity was used to measure PanNET tumoroid growth in the presence of HH pathway agonists or ( D ) inhibitors of the canonical and ( E ) non-canonical HH signaling pathways. Fluorescence signal is compared to DMSO vehicle control. ( n = 3 replicates in two unique mouse PanNET tumoroid lines). ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test. F BrdU incorporation was used to measure tumoroid proliferation in GFAP ΔMen1 and Sox10 ΔMen1 PanNET tumoroids after 72 h treatment. ( n = 4). ** = p < 0.01, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test. G Relative fold-change in Chga mRNA levels in PanNET tumoroids following 72 h treatment. ( n = 5). H Western blot analysis of HH pathway proteins in PanNET tumoroids after 72 h treatment. I Quantitation of SHH protein expression normalized to beta-actin and DMSO vehicle control from the western blot analysis in panel ( H ). ( n = 3). J Western blot analysis and ( K ) associated quantitation of phosphorylated and total ERK and AKT growth pathways in PanNET tumoroids after 72 h treatment. ( n = 3). * = p < 0.05, ** = p < 0.01, *** = p < 0.001 by Kruskal-Wallis test

Article Snippet: Drug compounds used in the studies include: human recombinant SHH N-terminal peptide (R&D Systems, Cat# 1845-GMP), SAG (Tocris, Cat# 4366), vismodegib (Tocris, Cat# 7710), sonidegib (Tocris, Cat# 7826), GANT61 (Tocris, Cat# 3191), and itraconazole (Tocris, Cat# 5981).

Techniques: Fluorescence, Protein-Protein interactions, Control, BrdU Incorporation Assay, Western Blot, Quantitation Assay, Expressing

Journal: Molecular Cancer

Article Title: Hedgehog signaling drives glial cell plasticity and oncogenic reprogramming in gastroenteropancreatic neuroendocrine neoplasms

doi: 10.1186/s12943-026-02611-y

Figure Lengend Snippet: GFAP ΔMen1 DNETs and jejunal NETs are sensitive to HH pathway activation and inhibition. A Phase contrast and ( B ) fluorescence images of DNET tumoroids from a GFAP-Cre; Men1 FL/FL ; LSL-tdTomato mouse. Tumoroids were imaged after 72 h exposure to: the HH agonists SHH-N (100 ng/mL) and SAG (10 nM); inhibitors of the canonical HH signaling pathway vismodegib (20 µM) and sonidegib (10 nM); or inhibitors of the GLI1/2 effectors GANT61 (10 µM) and itraconazole (ITZ, 1 µM). C TdTomato fluorescence intensity was used to measure DNET tumoroid growth in the presence of HH pathway inhibitors. Fluorescence signal is compared to DMSO vehicle control. ( n = 3 replicates using one mouse DNET tumoroid line). ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test. D Relative BrdU incorporation in a second GFAP ΔMen1 DNET tumoroid line and ( E ) a jejunal tumoroid line (J-NET) after 72 h treatment. ( n = 4). ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test. F Western blot analysis of SHH, ERK, and AKT growth pathways in J-NET tumoroids after 72 h treatment. G Western blot quantitation of SHH and ( H ) phosphorylated and total ERK and AKT proteins normalized to GAPDH and DMSO vehicle control. ( n = 3). * = p < 0.05 by Kruskal-Wallis test. I Crystal violet staining of mouse STC-1 SI-NET cells after 48 h treatment with agonists and inhibitors of the HH signaling pathway. J BrdU incorporation in STC-1 cells after 48 h treatment with HH agonists SHH-N and SAG, ( K ) SMO inhibitors vismodegib and sonidegib, and ( L ) inhibitors of GLI1/2 signaling. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test

Article Snippet: Drug compounds used in the studies include: human recombinant SHH N-terminal peptide (R&D Systems, Cat# 1845-GMP), SAG (Tocris, Cat# 4366), vismodegib (Tocris, Cat# 7710), sonidegib (Tocris, Cat# 7826), GANT61 (Tocris, Cat# 3191), and itraconazole (Tocris, Cat# 5981).

Techniques: Activation Assay, Inhibition, Fluorescence, Control, BrdU Incorporation Assay, Western Blot, Quantitation Assay, Staining

Journal: Cellular signalling

Article Title: Coordinated D -cyclin/Foxd1 activation drives mitogenic activity of the Sonic Hedgehog signaling pathway

doi: 10.1016/j.cellsig.2017.12.007

Figure Lengend Snippet: Canonical Shh signaling drives proliferation in iMEFs. (A) Wild-type (WT) and Gli2−/−3−/− iMEFs were cultured ± SHH ligand and ± the SMO antagonist vismodegib (Vis). Cell number is shown relative to vehicle-treated cells. SHH increased proliferation in WT iMEFs, but not in Gli2−/−3−/− iMEFs. This increase in proliferation was blocked by the addition of vismodegib (n = 5). (B) Histogram of fluorescence intensity of cells treated with the cell tracing reagent CellTrace™ Far Red and analyzed by flow cytometry. SHH treatment resulted in a leftward shift in fluorescence compared to vehicle-treated cells, indicating an increase in cellular divisions (n = 1). Significance: * = p < 0.05. Error bars show SEM.

Article Snippet: The SMO inhibitor, vismodegib, was purchased from LC Laboratories (CAS No 879085–55–9) and dissolved in DMSO.

Techniques: Cell Culture, Fluorescence, Flow Cytometry

Journal: Cellular signalling

Article Title: Coordinated D -cyclin/Foxd1 activation drives mitogenic activity of the Sonic Hedgehog signaling pathway

doi: 10.1016/j.cellsig.2017.12.007

Figure Lengend Snippet: Foxd1 is a target of canonical Sonic Hedgehog signaling. (A) Gene expression was determined for WT iMEFs cultured ± SHH ligand and ± the SMO antagonist vismodegib (Vis). Expression changes are shown as a fold change from vehicle-treated cells. SHH treatment resulted in an increase in Gli1 and Foxd1 expression, which was blocked by the addition of vismodegib (n = 6). (B) Expression of Gli1 and Foxd1 is increased in Ptch1−/− iMEFs relative to wild-type (inset) and significantly reduced by vismodegib treatment. (C) Expression of Gli1 and Foxd1 is increased in iMEFs overexpressing a constitutively active form of human Smoothened (SMOM2). Expression changes are shown as a fold change from cell overexpressing GFP (n = 5). Insert shows SMO expression. (D) Expression of Gli1 and Foxd1 is not significantly changed in Gli2−/−3−/− iMEFs treated with SHH ligand (n = 4). Significance: * = p < 0.05, ** = p < 0.01, *** = p < 0.001. Error bars show SEM.

Article Snippet: The SMO inhibitor, vismodegib, was purchased from LC Laboratories (CAS No 879085–55–9) and dissolved in DMSO.

Techniques: Gene Expression, Cell Culture, Expressing

Journal: Cellular signalling

Article Title: Coordinated D -cyclin/Foxd1 activation drives mitogenic activity of the Sonic Hedgehog signaling pathway

doi: 10.1016/j.cellsig.2017.12.007

Figure Lengend Snippet: SHH suppresses Cdkn1c expression. (A) Gene expression was determined for wild-type (WT) iMEFs cultured ± SHH ligand and ± the SMO antagonist vismodegib (Vis). Expression changes are shown as a fold change from vehicle-treated cells. SHH treatment resulted in an increase in Ccnd1 and a decrease in Cdkn1c expression, both of which were blocked by the addition of vismodegib. Expression changes are shown as a fold change from vehicle-treated cells. (n = 9). (B) Expression of Ccnd1 is increased in Ptch1−/− iMEFs relative to WT (inset). Vismodegib treatment Ptch1−/− iMEFs results in decreased Ccnd1 expression and increased Cdkn1c expression (C) Expression of Gli1 and Foxd1 is not significantly changed in Gli2−/− 3−/− iMEFs treated with SHH ligand (n = 4). (C) In iMEFs overexpressing a constitutively active form of human Smoothened (SMOM2), expression of Ccnd1 was increased while Cdkn1c expression was decreased. Expression changes are shown as a fold change from cells overexpressing GFP (n = 5). (D) To determine the temporal sequence of gene expression changes, WT cells were harvested at 6, 12, 24, 48, and 72 h after treatment ± SHH ligand. Gli1 expression (white arrow) is significantly increased by 6 h followed by increased Foxd1 expression (black arrow) by 24 h, and Cdkn1c expression (gray arrow) is not significantly decreased until 48 h post-treatment. Expression changes are shown as a fold change from vehicle-treated cells at respective time points (n = 5). Significance: * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001. Error bars show SEM.

Article Snippet: The SMO inhibitor, vismodegib, was purchased from LC Laboratories (CAS No 879085–55–9) and dissolved in DMSO.

Techniques: Expressing, Gene Expression, Cell Culture, Sequencing

Journal: The European Journal of Neuroscience

Article Title: Brain‐derived neurotrophic factor overexpression in taste buds diminishes chemotherapy induced taste loss

doi: 10.1111/ejn.15799

Figure Lengend Snippet: Taste perception is preserved in BDNF overexpressing mice following vismodegib treatment. There was a significantly higher taste preference in vehicle‐treated GB739 and GB759 transgenic mice compared to vehicle‐treated wild‐type controls. Vismodegib‐treated wild‐type mice showed a significant decrease in taste preference. Vismodegib‐treated GB759 mice presented a significantly higher preference for the sucrose solution compared to vismodegib‐treated wild‐type mice. Although there was a higher preference in vismodegib‐treated GB739 mice compared to their wild‐type mice counterparts, they did not reach the level of significance, but there is a trend. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: GB739, GB759 and wild‐type controls ( n = 15/genotype) were administered vismodegib (LC Laboratories, Woburn, MA; 30 mg/kg) orally in almond butter (Jif® Natural Creamy Almond Butter Spread) on a glass petri dish for 10 weeks.

Techniques: Transgenic Assay

Journal: The European Journal of Neuroscience

Article Title: Brain‐derived neurotrophic factor overexpression in taste buds diminishes chemotherapy induced taste loss

doi: 10.1111/ejn.15799

Figure Lengend Snippet: Overexpression of BDNF in taste buds prevents the decrease of taste bud size in circumvallate papillae. Vehicle‐treated GB mice had significantly larger taste buds than wild‐type mice. Additionally, vismodegib‐treated GB mice had larger taste buds compared to both vehicle‐ and vismodegib‐treated wild‐type mice. * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: GB739, GB759 and wild‐type controls ( n = 15/genotype) were administered vismodegib (LC Laboratories, Woburn, MA; 30 mg/kg) orally in almond butter (Jif® Natural Creamy Almond Butter Spread) on a glass petri dish for 10 weeks.

Techniques: Over Expression

Journal: The European Journal of Neuroscience

Article Title: Brain‐derived neurotrophic factor overexpression in taste buds diminishes chemotherapy induced taste loss

doi: 10.1111/ejn.15799

Figure Lengend Snippet: BDNF overexpression in taste buds preserves NCAM labelling. NCAM innervation was richer in taste buds of vehicle‐treated GB759 mice (b) compared to vehicle‐treated C57 (a) and preserved in vismodegib‐treated GB759 mice (d) compared to vismodegib‐treated C57 mice (c). The NCAM fluorescence intensity was significantly higher in both vismodegib‐ and vehicle‐treated GB759 mice compared to their wild‐type counterparts (e). There was no significant decrease in NCAM fluorescence intensity in vismodegib‐treated GB759, whereas there was a significant decrease in vismodegib‐treated wild‐type mice. Scale bar = 100 μm. ** p < 0.01; ** p < 0.01; **** p < 0.0001. NCAM, neural cell adhesion molecule 1

Article Snippet: GB739, GB759 and wild‐type controls ( n = 15/genotype) were administered vismodegib (LC Laboratories, Woburn, MA; 30 mg/kg) orally in almond butter (Jif® Natural Creamy Almond Butter Spread) on a glass petri dish for 10 weeks.

Techniques: Over Expression, Fluorescence

Journal: The European Journal of Neuroscience

Article Title: Brain‐derived neurotrophic factor overexpression in taste buds diminishes chemotherapy induced taste loss

doi: 10.1111/ejn.15799

Figure Lengend Snippet: BDNF overexpression protects P2X3 labelling of taste buds and gustatory innervation. IHC with P2X3 antibodies showed preserved innervation inside the taste buds (d, thick arrow) and in the subepithelial nerve plexus of the circumvallate papillae of vismodegib‐treated GB759 (d, thin arrow) compared to vismodegib‐treated wild‐type mice (c). Both vismodegib‐treated (d) and vehicle‐treated (b) GB759 mice showed stronger labeling compared to their wild‐type controls (a,c) Because of the orientation of the section, the subepithelial nerve plexus is not visible in vehicle‐treated GB759mice (b). There was a significant decrease in P2X3 fluorescence intensity in vismodegib‐treated wild‐type mice (e), whereas there was no significant decrease in vismodegib‐treated GB759 mice. Scale bar = 50 µm. * p < 0.05; *** p < 0.001. IHC, immunohistochemistry

Article Snippet: GB739, GB759 and wild‐type controls ( n = 15/genotype) were administered vismodegib (LC Laboratories, Woburn, MA; 30 mg/kg) orally in almond butter (Jif® Natural Creamy Almond Butter Spread) on a glass petri dish for 10 weeks.

Techniques: Over Expression, Labeling, Fluorescence, Immunohistochemistry

Journal: The European Journal of Neuroscience

Article Title: Brain‐derived neurotrophic factor overexpression in taste buds diminishes chemotherapy induced taste loss

doi: 10.1111/ejn.15799

Figure Lengend Snippet: Taste bud‐specific overexpression of BDNF counteracts the decrease of Shh‐like immunoreactive cells. Shh and Troma‐1 labelling were examined using IHC in C57 and GB circumvallate papillae sections. Superimposition of Shh (red) and Troma‐1 (green) resulted in yellow (co‐localisation). (a, c, e and g) IHC with Shh, and (b, d, f and h) double staining with Shh and Troma‐1. (c and d) Shh and Troma‐1 expressions are highest in vehicle‐treated GB739. (e and f) Shh and Troma‐1 labelling is lower in vismodegib‐treated C57 mice than in (g and h) vismodegib‐treated GB739 mice. (i) Shh fluorescence intensity was significantly higher in vismodegib‐treated GB739 mice compared to vismodegib‐treated wild‐type mice. Vismodegib‐treated wild‐type mice showed a significantly lower Shh fluorescence intensity than vehicle‐treated wild‐type mice. Scale bar = 100 μm. * p < 0.05; ** p < 0.01. IHC, immunohistochemistry; GB, Gustducin‐BDNF; BDNF, brain‐derived neurotrophic factor

Article Snippet: GB739, GB759 and wild‐type controls ( n = 15/genotype) were administered vismodegib (LC Laboratories, Woburn, MA; 30 mg/kg) orally in almond butter (Jif® Natural Creamy Almond Butter Spread) on a glass petri dish for 10 weeks.

Techniques: Over Expression, Double Staining, Fluorescence, Immunohistochemistry, Derivative Assay

Journal: The European Journal of Neuroscience

Article Title: Brain‐derived neurotrophic factor overexpression in taste buds diminishes chemotherapy induced taste loss

doi: 10.1111/ejn.15799

Figure Lengend Snippet: Reduced Ki‐67 and Gustducin labelling in wild‐type vallate taste cells compared to GB mice. (a, d, g and j) Double IHC was performed with Ki‐67 and (b, e, h and k) with Gustducin (c, f, i and l). (d) Ki67 (green) showed stronger labelling in GB759 vehicle‐treated mice compared to C57 vehicle‐treated mice (a). (g) Vismodegib‐treated C57 mice had a robust decrease in Ki67 labelling, whereas the decrease was less pronounced in GB759 drug‐treated mice (j). The same trend was observed with Gustducin (red). (e) Vehicle‐treated GB759 mice had a stronger Gustducin labelling compared to vehicle‐treated C57 mice (b). Gustducin labelling in vismodegib‐treated wild‐type mice (h) was weaker compared to vismodegib‐treated GB759 mice (k). (m and n) There was no significant Ki67, and Gustducin fluorescence intensity decrease in vismodegib‐treated GB759 mice, whereas vismodegib‐treated wild‐type mice showed a significant decrease. Scale bar = 100 μm. * p < 0.05; ** p < 0.01; **** p < 0.0001. IHC, immunohistochemistry; GB, Gustducin‐BDNF; BDNF, brain‐derived neurotrophic factor

Article Snippet: GB739, GB759 and wild‐type controls ( n = 15/genotype) were administered vismodegib (LC Laboratories, Woburn, MA; 30 mg/kg) orally in almond butter (Jif® Natural Creamy Almond Butter Spread) on a glass petri dish for 10 weeks.

Techniques: Fluorescence, Immunohistochemistry, Derivative Assay

Journal: The European Journal of Neuroscience

Article Title: Brain‐derived neurotrophic factor overexpression in taste buds diminishes chemotherapy induced taste loss

doi: 10.1111/ejn.15799

Figure Lengend Snippet: Preserved T1R3 labelling in vismodegib‐treated GB mice compared to wild‐type mice. IHC of circumvallate papillae showed stronger T1R3 labelling in vehicle‐treated GB circumvallate taste buds (b) compared to wild‐type mice (a). Vismodegib‐treated GB759 showed less decrease of T1R3 signals (d) compared to wild‐type vismodegib‐treated mice (c). (e) There was no significant decrease in T1R3 fluorescence intensity in vismodegib‐treated GB759 mice, whereas there was a significant decrease in vismodegib‐treated wild‐type mice. The fluorescence intensity in vismodegib‐treated GB759 mice was significantly higher compared to vismodegib‐treated wild‐type mice. Scale bar = 100 μm. ** p < 0.01; *** p < 0.0001. IHC, immunohistochemistry; GB, Gustducin‐BDNF; BDNF, brain‐derived neurotrophic factor

Article Snippet: GB739, GB759 and wild‐type controls ( n = 15/genotype) were administered vismodegib (LC Laboratories, Woburn, MA; 30 mg/kg) orally in almond butter (Jif® Natural Creamy Almond Butter Spread) on a glass petri dish for 10 weeks.

Techniques: Fluorescence, Immunohistochemistry, Derivative Assay

Journal: Nature Communications

Article Title: Identification of recurrent FHL2-GLI2 oncogenic fusion in sclerosing stromal tumors of the ovary

doi: 10.1038/s41467-019-13806-x

Figure Lengend Snippet: a Cell titer blue proliferation assay of immortalized mesenchymal stem cells (MSCs), HEK-293, medulloblastoma (DAOY) and human basal cell carcinoma (BCC) cells stably expressing empty vector (control), wild-type FHL2 (FHL2), wild-type GLI2 (GLI2), truncated GLI2 (tGLI2) or FHL2-GLI2 treated with 250 nM Vismodegib or vehicle control (DMSO). b Representative images of colony formation assay of MSC, HEK-293, DAOY, and BCC cells stably expressing control, FHL2, GLI2, tGLI2, or FHL2-GLI2 treated with Vismodegib 500 nM or vehicle control (DMSO). Scale bars, 5 mm. Quantification of the number of colonies/well compared to control (bottom). c Wound healing assay of MSC, HEK-293, DAOY, and BCC cells stably expressing control, FHL2, GLI2, tGLI2, or FHL2-GLI2 treated with 250 nM Vismodegib or vehicle control (DMSO). The migratory effect/wound area was assessed at 0 and 24 h and quantified compared to DMSO (bottom). Vismo, Vismodegib. Scale bars, 500 μm. In a – c , data are representative of at least three independent experiments. Error bars, s.d. of mean; n.s., not significant; * P < 0.05, ** P < 0.01, *** P < 0.001; two-tailed unpaired t -test.

Article Snippet: The smoothened inhibitors Cyclopamine (Selleckchem, S1146), Vismodegib (Cellagen tech, C4044–5) and the GLI inhibitor GANT61 (Tocris, 3191) were resuspended in DMSO and used in in vitro proliferation, colony formation and scratch wound healing assays at 5–10 μM, 250–500 nM, and 5–10 μM, respectively.

Techniques: Proliferation Assay, Stable Transfection, Expressing, Plasmid Preparation, Control, Colony Assay, Wound Healing Assay, Two Tailed Test