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    Name:
    GLI 1 Lentiviral Activation Particles
    Description:
    CRISPR Cas9 KO Plasmids consists of GLI 1 specific 20 nt guide RNA sequences derived from the GeCKO v2 library For CRISPR gene knockout gRNA sequences direct the Cas9 protein to induce a site specific double strand break DSB in the genomic DNA Gene knockdown or activation can be assayed using GLI 1 Antibody C 1 sc 515751 All products are provided as transfection ready purified plasmid DNA except the Lentiviral Activation Particles which have been prepackaged as Lentiviral Particles for use with hard to transfect cells
    Catalog Number:
    SC-400266-LAC
    Price:
    None
    Category:
    Gene Editing Transcription Regulator GLI 1 CRISPR Plasmids GLI 1 CRISPR Plasmids h
    Buy from Supplier


    Structured Review

    Santa Cruz Biotechnology gli1
    Tumor cells competent for Hh signaling and OPN expression are efficient at inducing osteoblast differentiation. Stable silencing of OPN (OPNi) or <t>GLI1</t> (KD2 and KO1) from SUM1315 and MDA-MB-435 tumor cells significantly reduces the expression of ( A ) BSP [Relative to SUM1315, SUM1315-OPNi (∧p = 0.008) and KD2 (∧p = 0.0004) show lower BSP; Relative to MDA-MB-435, 435-OPNi and KO1 have decreased BSP (∧p
    CRISPR Cas9 KO Plasmids consists of GLI 1 specific 20 nt guide RNA sequences derived from the GeCKO v2 library For CRISPR gene knockout gRNA sequences direct the Cas9 protein to induce a site specific double strand break DSB in the genomic DNA Gene knockdown or activation can be assayed using GLI 1 Antibody C 1 sc 515751 All products are provided as transfection ready purified plasmid DNA except the Lentiviral Activation Particles which have been prepackaged as Lentiviral Particles for use with hard to transfect cells
    https://www.bioz.com/result/gli1/product/Santa Cruz Biotechnology
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gli1 - by Bioz Stars, 2021-09
    99/100 stars

    Images

    1) Product Images from "Hedgehog Signaling in Tumor Cells Facilitates Osteoblast-Enhanced Osteolytic Metastases"

    Article Title: Hedgehog Signaling in Tumor Cells Facilitates Osteoblast-Enhanced Osteolytic Metastases

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0034374

    Tumor cells competent for Hh signaling and OPN expression are efficient at inducing osteoblast differentiation. Stable silencing of OPN (OPNi) or GLI1 (KD2 and KO1) from SUM1315 and MDA-MB-435 tumor cells significantly reduces the expression of ( A ) BSP [Relative to SUM1315, SUM1315-OPNi (∧p = 0.008) and KD2 (∧p = 0.0004) show lower BSP; Relative to MDA-MB-435, 435-OPNi and KO1 have decreased BSP (∧p
    Figure Legend Snippet: Tumor cells competent for Hh signaling and OPN expression are efficient at inducing osteoblast differentiation. Stable silencing of OPN (OPNi) or GLI1 (KD2 and KO1) from SUM1315 and MDA-MB-435 tumor cells significantly reduces the expression of ( A ) BSP [Relative to SUM1315, SUM1315-OPNi (∧p = 0.008) and KD2 (∧p = 0.0004) show lower BSP; Relative to MDA-MB-435, 435-OPNi and KO1 have decreased BSP (∧p

    Techniques Used: Expressing, Multiple Displacement Amplification

    Active Hh signaling in the tumor cells is vital to their ability to cause osteolysis. MDA-MB-435 tumor cells were injected into the left cardiac ventricle of athymic mice; mice were euthanized 4–6 weeks later and radiographically imaged and assessed for osteolysis at the tibio-femoral junction. As represented in the Table, cells that were silenced for GLI1 expression showed an attenuated ability of osteolysis. The percent incidence of osteolysis is depicted in the adjacent graph.
    Figure Legend Snippet: Active Hh signaling in the tumor cells is vital to their ability to cause osteolysis. MDA-MB-435 tumor cells were injected into the left cardiac ventricle of athymic mice; mice were euthanized 4–6 weeks later and radiographically imaged and assessed for osteolysis at the tibio-femoral junction. As represented in the Table, cells that were silenced for GLI1 expression showed an attenuated ability of osteolysis. The percent incidence of osteolysis is depicted in the adjacent graph.

    Techniques Used: Multiple Displacement Amplification, Injection, Mouse Assay, Expressing

    The Hh pathway is activated in breast tumors. Breast cancer tissues (n = 75) and normal breast tissues (n = 9) were immunohistochemically stained for ( A ) IHH and ( B ) GLI1 expression. The intensity of cytoplasmic staining was quantitated with computer-assisted image analysis in a Dako ACIS III Image Analysis System (Glostrup, Denmark). Staining intensities were recorded and represented as a scatter plot. The staining intensities indicating expression levels of IHH and GLI1 were significantly greater (p
    Figure Legend Snippet: The Hh pathway is activated in breast tumors. Breast cancer tissues (n = 75) and normal breast tissues (n = 9) were immunohistochemically stained for ( A ) IHH and ( B ) GLI1 expression. The intensity of cytoplasmic staining was quantitated with computer-assisted image analysis in a Dako ACIS III Image Analysis System (Glostrup, Denmark). Staining intensities were recorded and represented as a scatter plot. The staining intensities indicating expression levels of IHH and GLI1 were significantly greater (p

    Techniques Used: Staining, Expressing

    2) Product Images from "Hedgehog/Gli promotes epithelial-mesenchymal transition in lung squamous cell carcinomas"

    Article Title: Hedgehog/Gli promotes epithelial-mesenchymal transition in lung squamous cell carcinomas

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/1756-9966-33-34

    Gli1 expression reversely correlates with E-Cadherin expression in lung SCC. (A) Expressions of Gli1 and E-Cadherin (E-Cad) in three representative tissue specimens in the UCSF cohort with Gli1 expression at a low level (upper panels) and high levels (middle and lower panels). (B) Expressions of Gli1, E-Cad and β-Catenin (β-Cat) in three representative tissue specimens in the Tianjin cohort with Gli1 expression at a low level (upper panels), a mixed expression pattern (middle panels) and a high level (lower panels). (C) Correlations between Gli1, EMT markers, and recurrence/metastasis. Statistical analysis was performed between Gli1 and E-Cad, Gli1 and β-Cat, Gli1 and recurrence/ metastasis. (D) Gli1 and E-Cad expression in four lung SCC cell lines by Western blots.
    Figure Legend Snippet: Gli1 expression reversely correlates with E-Cadherin expression in lung SCC. (A) Expressions of Gli1 and E-Cadherin (E-Cad) in three representative tissue specimens in the UCSF cohort with Gli1 expression at a low level (upper panels) and high levels (middle and lower panels). (B) Expressions of Gli1, E-Cad and β-Catenin (β-Cat) in three representative tissue specimens in the Tianjin cohort with Gli1 expression at a low level (upper panels), a mixed expression pattern (middle panels) and a high level (lower panels). (C) Correlations between Gli1, EMT markers, and recurrence/metastasis. Statistical analysis was performed between Gli1 and E-Cad, Gli1 and β-Cat, Gli1 and recurrence/ metastasis. (D) Gli1 and E-Cad expression in four lung SCC cell lines by Western blots.

    Techniques Used: Expressing, Western Blot

    Aberrant activation of the Shh pathway in lung SCC. (A) Representative protein expression of Shh, Smo, Ptch1 and Gli1 by IHC staining, scored as 0 (negative), 1 (mild positive), 2 (positive), and 3 (strong positive). (B) Expression distributions of Shh, Smo, Ptch1 and Gli1 in 177 patient tissue specimens in the Tianjin cohort. (C) Association between the expressions of Hh pathway components. Kendall’s tau-b statsitcs was used to determine the correlation between proteins. The correlation coefficient r and p values were presented in (C) . Kappa test was also performed with IHC scores of 1–3 grouped as “+”, 0 as “-”. Kappa test’s p values were labeled with*.
    Figure Legend Snippet: Aberrant activation of the Shh pathway in lung SCC. (A) Representative protein expression of Shh, Smo, Ptch1 and Gli1 by IHC staining, scored as 0 (negative), 1 (mild positive), 2 (positive), and 3 (strong positive). (B) Expression distributions of Shh, Smo, Ptch1 and Gli1 in 177 patient tissue specimens in the Tianjin cohort. (C) Association between the expressions of Hh pathway components. Kendall’s tau-b statsitcs was used to determine the correlation between proteins. The correlation coefficient r and p values were presented in (C) . Kappa test was also performed with IHC scores of 1–3 grouped as “+”, 0 as “-”. Kappa test’s p values were labeled with*.

    Techniques Used: Activation Assay, Expressing, Immunohistochemistry, Staining, Labeling

    3) Product Images from "MTOR promotes basal cell carcinoma growth through atypical PKC"

    Article Title: MTOR promotes basal cell carcinoma growth through atypical PKC

    Journal: bioRxiv

    doi: 10.1101/2020.08.24.264598

    mTor inhibition suppresses murine BCC growth and aPKC activity. A) Hematoxylin and eosin staining of dorsal back skin collected from DMSO- or Everolimus-treated Ptch1 fl/fl ; Gli1 - Cre ERT2 mice. Scale bar, 50 μm. B) Quantification of total tumor size per square area (n > 250 tumors from 5 mice). tx, treatment. C) Immunofluorescent staining of DMSO- or Everolimus-treated Ptch1 fl/fl ; Gli1 - Cre ERT2 mice for the indicated markers. Scale bar, 25 μm. D) Quantification of immunostains (n = 5 tumors from 3 mice). AU, arbitrary units. Error bars represent SEM. Significance was determined by unpaired two-tailed t test. *, p
    Figure Legend Snippet: mTor inhibition suppresses murine BCC growth and aPKC activity. A) Hematoxylin and eosin staining of dorsal back skin collected from DMSO- or Everolimus-treated Ptch1 fl/fl ; Gli1 - Cre ERT2 mice. Scale bar, 50 μm. B) Quantification of total tumor size per square area (n > 250 tumors from 5 mice). tx, treatment. C) Immunofluorescent staining of DMSO- or Everolimus-treated Ptch1 fl/fl ; Gli1 - Cre ERT2 mice for the indicated markers. Scale bar, 25 μm. D) Quantification of immunostains (n = 5 tumors from 3 mice). AU, arbitrary units. Error bars represent SEM. Significance was determined by unpaired two-tailed t test. *, p

    Techniques Used: Inhibition, Activity Assay, Staining, Mouse Assay, Two Tailed Test

    mTor inhibition suppresses BCC cell growth but not HH signaling. A) Gli1 mRNA levels of ASZ001 cells treated with DMSO or varying concentrations of Rapamycin, OSI-027, or Everolimus (n = 3 experiments). dR, delta reporter signal normalized to passive reference dye. B-D) MTT assay of ASZ001 cells treated with B) Everolimus, C) Rapamycin, or D) OSI-027 (n = 3 experiments). Abs, absorbance. Error bars represent SEM. Significance was determined by two-way ANOVA test. *, p
    Figure Legend Snippet: mTor inhibition suppresses BCC cell growth but not HH signaling. A) Gli1 mRNA levels of ASZ001 cells treated with DMSO or varying concentrations of Rapamycin, OSI-027, or Everolimus (n = 3 experiments). dR, delta reporter signal normalized to passive reference dye. B-D) MTT assay of ASZ001 cells treated with B) Everolimus, C) Rapamycin, or D) OSI-027 (n = 3 experiments). Abs, absorbance. Error bars represent SEM. Significance was determined by two-way ANOVA test. *, p

    Techniques Used: Inhibition, MTT Assay

    MTOR is overexpressed in human and mouse BCC. A) Immunofluorescent staining of indicated markers in human normal epidermis and nodular BCC. Scale bar, 50 μm. B) Quantification of MTOR immunostain intensity (n = 5 different points of measurement from 4 individual samples). AU, arbitrary units. C) Immunofluorescent staining of indicated markers in normal epidermis, normal hair follicle, or BCC derived from Ptch1 fl/fl ; Gli1 - Cre ERT2 mice. Scale bar, 25 μm. D) Quantification of mTor immunostain (n = 5 different points of measurement from 5 mice). Error bars represent SEM. Significance was determined by unpaired two-tailed t test. *, p
    Figure Legend Snippet: MTOR is overexpressed in human and mouse BCC. A) Immunofluorescent staining of indicated markers in human normal epidermis and nodular BCC. Scale bar, 50 μm. B) Quantification of MTOR immunostain intensity (n = 5 different points of measurement from 4 individual samples). AU, arbitrary units. C) Immunofluorescent staining of indicated markers in normal epidermis, normal hair follicle, or BCC derived from Ptch1 fl/fl ; Gli1 - Cre ERT2 mice. Scale bar, 25 μm. D) Quantification of mTor immunostain (n = 5 different points of measurement from 5 mice). Error bars represent SEM. Significance was determined by unpaired two-tailed t test. *, p

    Techniques Used: Staining, Derivative Assay, Mouse Assay, Two Tailed Test

    4) Product Images from "The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *"

    Article Title: The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M109.021949

    The Hh pathway transcriptionally regulates OPN. A , levels of GLI1 and OPN are increased in primary cutaneous cancer and metastatic melanoma. Gene microarray analysis (utilizing a Human Genome U133 Plus 2.0 array from Affymetrix, Inc.) was used to compare
    Figure Legend Snippet: The Hh pathway transcriptionally regulates OPN. A , levels of GLI1 and OPN are increased in primary cutaneous cancer and metastatic melanoma. Gene microarray analysis (utilizing a Human Genome U133 Plus 2.0 array from Affymetrix, Inc.) was used to compare

    Techniques Used: Microarray

    Silencing endogenous GLI1 expression diminishes attributes of motility, invasion, migration, and proliferation and negatively impacts tumorigenicity and metastasis. A , abrogating GLI1 expression ( KO1 and KO4 ) reduced the ability of cells to move and fill
    Figure Legend Snippet: Silencing endogenous GLI1 expression diminishes attributes of motility, invasion, migration, and proliferation and negatively impacts tumorigenicity and metastasis. A , abrogating GLI1 expression ( KO1 and KO4 ) reduced the ability of cells to move and fill

    Techniques Used: Expressing, Migration

    The OPN promoter bears a GLI1-binding site that responds to GLI1. A , mutating the putative GLI1-binding site in the OPN promoter makes the OPN promoter insensitive to the effects of GLI1. The OPN promoter (+112 to −352 (pGL3-OPN-352)) was significantly
    Figure Legend Snippet: The OPN promoter bears a GLI1-binding site that responds to GLI1. A , mutating the putative GLI1-binding site in the OPN promoter makes the OPN promoter insensitive to the effects of GLI1. The OPN promoter (+112 to −352 (pGL3-OPN-352)) was significantly

    Techniques Used: Binding Assay

    Restoring the availability of OPN-initiated signaling in GLI1-silenced cells reinstates their motility and ability to migrate and invade. A , immunoblot representing the restored OPN in cells that have been stably knocked down for GLI1 (GLI1 KO; and consequently
    Figure Legend Snippet: Restoring the availability of OPN-initiated signaling in GLI1-silenced cells reinstates their motility and ability to migrate and invade. A , immunoblot representing the restored OPN in cells that have been stably knocked down for GLI1 (GLI1 KO; and consequently

    Techniques Used: Stable Transfection

    shRNA to GLI1 abrogates expression of OPN and brings about a partial reversal of EMT. A , MDA-MB-435 cells were stably transfected with shRNA to GLI1. Clones KO1 to KO4 were stably silenced for GLI1 and show notably reduced OPN expression. B , real time
    Figure Legend Snippet: shRNA to GLI1 abrogates expression of OPN and brings about a partial reversal of EMT. A , MDA-MB-435 cells were stably transfected with shRNA to GLI1. Clones KO1 to KO4 were stably silenced for GLI1 and show notably reduced OPN expression. B , real time

    Techniques Used: shRNA, Expressing, Multiple Displacement Amplification, Stable Transfection, Transfection, Clone Assay

    5) Product Images from "Trimeric G protein-CARMA1 axis links smoothened, the hedgehog receptor transducer, to NF-?B activation in diffuse large B-cell lymphoma"

    Article Title: Trimeric G protein-CARMA1 axis links smoothened, the hedgehog receptor transducer, to NF-?B activation in diffuse large B-cell lymphoma

    Journal: Blood

    doi: 10.1182/blood-2012-12-470153

    SMO activates NF-κB pathway independent of GLI1. (A) Silencing SMO in DOHH2 and HBL1 resulted in decrease of p -p65, (B) decrease of nuclear DNA binding activity of p65 and p50, and (C) decrease of total NF-κB luciferase activity. (D) Silencing
    Figure Legend Snippet: SMO activates NF-κB pathway independent of GLI1. (A) Silencing SMO in DOHH2 and HBL1 resulted in decrease of p -p65, (B) decrease of nuclear DNA binding activity of p65 and p50, and (C) decrease of total NF-κB luciferase activity. (D) Silencing

    Techniques Used: Binding Assay, Activity Assay, Luciferase

    Hh and NF-κB pathways are positively correlated in DLBCL, and Hh pathway contributes to NF-κB activation in DLBCL. (A) GLI1 expression levels in 145 DLBCL tumor biopsies. (B) High GLI1 expression was more frequent in DLBCL of ABC type
    Figure Legend Snippet: Hh and NF-κB pathways are positively correlated in DLBCL, and Hh pathway contributes to NF-κB activation in DLBCL. (A) GLI1 expression levels in 145 DLBCL tumor biopsies. (B) High GLI1 expression was more frequent in DLBCL of ABC type

    Techniques Used: Activation Assay, Expressing

    6) Product Images from "Defining Causative Factors Contributing in the Activation of Hedgehog Signaling in Diffuse Large B-Cell Lymphoma"

    Article Title: Defining Causative Factors Contributing in the Activation of Hedgehog Signaling in Diffuse Large B-Cell Lymphoma

    Journal: Leukemia research

    doi: 10.1016/j.leukres.2012.06.014

    The activation status of PI3K and NF-kB modulates GLI1 and Hh levels in DLBCL cell lines
    Figure Legend Snippet: The activation status of PI3K and NF-kB modulates GLI1 and Hh levels in DLBCL cell lines

    Techniques Used: Activation Assay

    7) Product Images from "Defining Causative Factors Contributing in the Activation of Hedgehog Signaling in Diffuse Large B-Cell Lymphoma"

    Article Title: Defining Causative Factors Contributing in the Activation of Hedgehog Signaling in Diffuse Large B-Cell Lymphoma

    Journal: Leukemia research

    doi: 10.1016/j.leukres.2012.06.014

    The activation status of PI3K and NF-kB modulates GLI1 and Hh levels in DLBCL cell lines
    Figure Legend Snippet: The activation status of PI3K and NF-kB modulates GLI1 and Hh levels in DLBCL cell lines

    Techniques Used: Activation Assay

    8) Product Images from "Gli is activated and promotes epithelial-mesenchymal transition in human esophageal adenocarcinoma"

    Article Title: Gli is activated and promotes epithelial-mesenchymal transition in human esophageal adenocarcinoma

    Journal: Oncotarget

    doi: 10.18632/oncotarget.22856

    Gli inhibition and siRNA knockdown reduce EMT and cell cycle activity in EAC cell lines (A) Western blots of OE19 and OE33 cells, pretreated with either DMSO (control) or Gli-i (500 nmol/L). Expression of AKT, p-AKT, Snail, Slug, and p21 was examined, with GAPDH serving as a loading control. (B) Western blots of EMT and cell cycle signaling markers in (i) OE19 (Gli1, p-S6K1, ERK, p-ERK, E-cadherin, N-cadherin, Cyclin D1), and (ii) OE33 (Gli1, Gli2, m-TOR, E-cadherin, N-cadherin, β-catenin, Cyclin D1), with GAPDH as a control. Cells were pretreated with DMSO, N-Shh (0.5 mg/mL), Gli-i (500 nmol/L), or Gli-i (500 nmol/L) with N-Shh (0.5 mg/mL) stimulation prior to protein extraction. (C) Western blots of OE19 and OE33 cells, subjected to siRNA treatment. Cells were given control siRNA (100 nM) or Gli1 + Gli2 siRNA (100 nM each) for 48 hours prior to total protein extraction. Expression of Gli1, Gli2, E-cadherin, N-cadherin, and β-catenin was examined, with GAPDH serving as an internal control.
    Figure Legend Snippet: Gli inhibition and siRNA knockdown reduce EMT and cell cycle activity in EAC cell lines (A) Western blots of OE19 and OE33 cells, pretreated with either DMSO (control) or Gli-i (500 nmol/L). Expression of AKT, p-AKT, Snail, Slug, and p21 was examined, with GAPDH serving as a loading control. (B) Western blots of EMT and cell cycle signaling markers in (i) OE19 (Gli1, p-S6K1, ERK, p-ERK, E-cadherin, N-cadherin, Cyclin D1), and (ii) OE33 (Gli1, Gli2, m-TOR, E-cadherin, N-cadherin, β-catenin, Cyclin D1), with GAPDH as a control. Cells were pretreated with DMSO, N-Shh (0.5 mg/mL), Gli-i (500 nmol/L), or Gli-i (500 nmol/L) with N-Shh (0.5 mg/mL) stimulation prior to protein extraction. (C) Western blots of OE19 and OE33 cells, subjected to siRNA treatment. Cells were given control siRNA (100 nM) or Gli1 + Gli2 siRNA (100 nM each) for 48 hours prior to total protein extraction. Expression of Gli1, Gli2, E-cadherin, N-cadherin, and β-catenin was examined, with GAPDH serving as an internal control.

    Techniques Used: Inhibition, Activity Assay, Western Blot, Expressing, Protein Extraction

    Gli1/2, EMT, and AKT pathway markers are upregulated in EAC tissue samples (A) Western blots of Gli1 and Gli2 protein expression in 24 matched pairs of esophageal tumor (T) and normal (N) tissues. GAPDH served as a loading control; results of 6 numbered, representative sample pairs are shown here. EMT markers (E-cadherin, N-cadherin, Vimentin, and β-catenin) were also examined. (B) Western blots of AKT pathway protein (p-MTOR, p-AKT, and p-S6K1) expression in 24 matched pairs of esophageal T and N tissues. GAPDH served as a loading control; results of 6 representative sample pairs are shown.
    Figure Legend Snippet: Gli1/2, EMT, and AKT pathway markers are upregulated in EAC tissue samples (A) Western blots of Gli1 and Gli2 protein expression in 24 matched pairs of esophageal tumor (T) and normal (N) tissues. GAPDH served as a loading control; results of 6 numbered, representative sample pairs are shown here. EMT markers (E-cadherin, N-cadherin, Vimentin, and β-catenin) were also examined. (B) Western blots of AKT pathway protein (p-MTOR, p-AKT, and p-S6K1) expression in 24 matched pairs of esophageal T and N tissues. GAPDH served as a loading control; results of 6 representative sample pairs are shown.

    Techniques Used: Western Blot, Expressing

    9) Product Images from "Trichostatin A promotes GLI1 degradation and P21 expression in multiple myeloma cells"

    Article Title: Trichostatin A promotes GLI1 degradation and P21 expression in multiple myeloma cells

    Journal: Cancer Management and Research

    doi: 10.2147/CMAR.S167330

    TSA enhances the degradation of GLI1. Notes: (A) GLI1 protein degradation in RPMI8226 and MM.1S cells treated with 5 μM TSA for 24 h and then 20 μM cycloheximide for up to 4 h. (B) Accumulation of GLI1 protein in RPMI8226 and MM.1S cells treated with 5 μM TSA and 10 μM MG132 for 48 h. (C) Immunoprecipitation assay showed the acetylation status of GLI1 in TSA-treated RPMI8226 and MM.1S cells. (D) Ubiquitination status of GLI1 in TSA-treated RPMI8226 and MM.1S cells were detected with immunoprecipitation. Representative images are from at least three independent experiments. Abbreviations: TSA, trichostatin A; CHX, cycloheximide; Con, control; Veh, vehicle; IB, immunoblot; IP: Immunoprecipitation.
    Figure Legend Snippet: TSA enhances the degradation of GLI1. Notes: (A) GLI1 protein degradation in RPMI8226 and MM.1S cells treated with 5 μM TSA for 24 h and then 20 μM cycloheximide for up to 4 h. (B) Accumulation of GLI1 protein in RPMI8226 and MM.1S cells treated with 5 μM TSA and 10 μM MG132 for 48 h. (C) Immunoprecipitation assay showed the acetylation status of GLI1 in TSA-treated RPMI8226 and MM.1S cells. (D) Ubiquitination status of GLI1 in TSA-treated RPMI8226 and MM.1S cells were detected with immunoprecipitation. Representative images are from at least three independent experiments. Abbreviations: TSA, trichostatin A; CHX, cycloheximide; Con, control; Veh, vehicle; IB, immunoblot; IP: Immunoprecipitation.

    Techniques Used: Immunoprecipitation

    TSA reduces GLI1 and downstream genes of hedgehog signaling. Notes: (A) The time- and dose-dependent downregulation of GLI1 after TSA treatment. Representative images from at least three independent experiments are shown. (B) The effect of TSA treatment for 48 h on GLI1 expression in mRNA level. (C) Luciferase assay showed the activity of the 8×GLI luciferase reporter responsive to increasing dosages of TSA. (D) Immunofluorescence showed the subcellular localization of GLI1 with or without TSA treatment. Magnification: 200×. Representative images are taken over 10–50 fields of view. Arrows, cells with GLI1 translocalized into the cytoplasm. (E) Statistical analysis of GLI1 localization in the nucleus of RPMI8226 and MM.1S cells treated with TSA for 24 h. (F) Representative images of Western blot analysis are shown to detect the subcellular distribution of GLI1. Histone H3 was used as an internal control for nuclear, and actin was used as an internal control for cytoplasm. (G) mRNA levels of c-MYC and SURVIVIN were detected by real-time PCR in MM cells treated with 5 μM TSA for 48 h. The median expression value of control group is normalized to 1. Data represent mean ± SD from three independent experiments ( # P
    Figure Legend Snippet: TSA reduces GLI1 and downstream genes of hedgehog signaling. Notes: (A) The time- and dose-dependent downregulation of GLI1 after TSA treatment. Representative images from at least three independent experiments are shown. (B) The effect of TSA treatment for 48 h on GLI1 expression in mRNA level. (C) Luciferase assay showed the activity of the 8×GLI luciferase reporter responsive to increasing dosages of TSA. (D) Immunofluorescence showed the subcellular localization of GLI1 with or without TSA treatment. Magnification: 200×. Representative images are taken over 10–50 fields of view. Arrows, cells with GLI1 translocalized into the cytoplasm. (E) Statistical analysis of GLI1 localization in the nucleus of RPMI8226 and MM.1S cells treated with TSA for 24 h. (F) Representative images of Western blot analysis are shown to detect the subcellular distribution of GLI1. Histone H3 was used as an internal control for nuclear, and actin was used as an internal control for cytoplasm. (G) mRNA levels of c-MYC and SURVIVIN were detected by real-time PCR in MM cells treated with 5 μM TSA for 48 h. The median expression value of control group is normalized to 1. Data represent mean ± SD from three independent experiments ( # P

    Techniques Used: Expressing, Luciferase, Activity Assay, Immunofluorescence, Western Blot, Real-time Polymerase Chain Reaction

    GLI1 overexpression rescues TSA-induced MM cell apoptosis. Notes: (A) Western blot assay confirmed the GLI1 protein in MM cells transfected with GLI1 expressing (OE) plasmid. (B) TSA treatment induced cleavage of PARP-1 as indicative of apoptosis in the GLI1 overexpressing MM cells compared with the vector controls. (C) Representative flow cytometry analysis of MM cells apoptosis treated with TSA for 48 h using Annexin V-FITC/PI staining. (D) Statistical analysis of three similar experiments for the MM cell apoptosis ( # P
    Figure Legend Snippet: GLI1 overexpression rescues TSA-induced MM cell apoptosis. Notes: (A) Western blot assay confirmed the GLI1 protein in MM cells transfected with GLI1 expressing (OE) plasmid. (B) TSA treatment induced cleavage of PARP-1 as indicative of apoptosis in the GLI1 overexpressing MM cells compared with the vector controls. (C) Representative flow cytometry analysis of MM cells apoptosis treated with TSA for 48 h using Annexin V-FITC/PI staining. (D) Statistical analysis of three similar experiments for the MM cell apoptosis ( # P

    Techniques Used: Over Expression, Western Blot, Transfection, Expressing, Plasmid Preparation, Flow Cytometry, Cytometry, Staining

    p21 induced by TSA represses the transcription of GLI1 . Notes: (A) The mRNA expression of GLI1 was detected by real-time PCR in MM cells transfected with p300 or p21 expressing (OE) plasmids compared with the vector control (vector). (B) GLI1 protein level in MM cells transfected with an increasing amount of p21 plasmid. ( C ) Half-life of GLI1 protein in RPMI8226 and MM.1S cells transfected with p21 or control plasmids was detected by Western blot. (D) RPMI8226 and MM.1S cells were transiently transfected with p21-specific or control siRNAs. Twenty-four hours post-transfection, cells were subjected for TSA treatment for another 24 h, and then, GLI1 protein was evaluated by Western blot. (E) Overexpression of p21 in RPMI8226 and MM.1S cells enhanced TSA-induced GLI1 repression. Abbreviations: TSA, trichostatin A; Veh, vehicle; MM, multiple myeloma.
    Figure Legend Snippet: p21 induced by TSA represses the transcription of GLI1 . Notes: (A) The mRNA expression of GLI1 was detected by real-time PCR in MM cells transfected with p300 or p21 expressing (OE) plasmids compared with the vector control (vector). (B) GLI1 protein level in MM cells transfected with an increasing amount of p21 plasmid. ( C ) Half-life of GLI1 protein in RPMI8226 and MM.1S cells transfected with p21 or control plasmids was detected by Western blot. (D) RPMI8226 and MM.1S cells were transiently transfected with p21-specific or control siRNAs. Twenty-four hours post-transfection, cells were subjected for TSA treatment for another 24 h, and then, GLI1 protein was evaluated by Western blot. (E) Overexpression of p21 in RPMI8226 and MM.1S cells enhanced TSA-induced GLI1 repression. Abbreviations: TSA, trichostatin A; Veh, vehicle; MM, multiple myeloma.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Western Blot, Over Expression

    10) Product Images from "Trimeric G protein-CARMA1 axis links smoothened, the hedgehog receptor transducer, to NF-?B activation in diffuse large B-cell lymphoma"

    Article Title: Trimeric G protein-CARMA1 axis links smoothened, the hedgehog receptor transducer, to NF-?B activation in diffuse large B-cell lymphoma

    Journal: Blood

    doi: 10.1182/blood-2012-12-470153

    SMO activates NF-κB pathway independent of GLI1. (A) Silencing SMO in DOHH2 and HBL1 resulted in decrease of p -p65, (B) decrease of nuclear DNA binding activity of p65 and p50, and (C) decrease of total NF-κB luciferase activity. (D) Silencing
    Figure Legend Snippet: SMO activates NF-κB pathway independent of GLI1. (A) Silencing SMO in DOHH2 and HBL1 resulted in decrease of p -p65, (B) decrease of nuclear DNA binding activity of p65 and p50, and (C) decrease of total NF-κB luciferase activity. (D) Silencing

    Techniques Used: Binding Assay, Activity Assay, Luciferase

    Hh and NF-κB pathways are positively correlated in DLBCL, and Hh pathway contributes to NF-κB activation in DLBCL. (A) GLI1 expression levels in 145 DLBCL tumor biopsies. (B) High GLI1 expression was more frequent in DLBCL of ABC type
    Figure Legend Snippet: Hh and NF-κB pathways are positively correlated in DLBCL, and Hh pathway contributes to NF-κB activation in DLBCL. (A) GLI1 expression levels in 145 DLBCL tumor biopsies. (B) High GLI1 expression was more frequent in DLBCL of ABC type

    Techniques Used: Activation Assay, Expressing

    11) Product Images from "Hedgehog/Gli promotes epithelial-mesenchymal transition in lung squamous cell carcinomas"

    Article Title: Hedgehog/Gli promotes epithelial-mesenchymal transition in lung squamous cell carcinomas

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/1756-9966-33-34

    Gli1 expression reversely correlates with E-Cadherin expression in lung SCC. (A) Expressions of Gli1 and E-Cadherin (E-Cad) in three representative tissue specimens in the UCSF cohort with Gli1 expression at a low level (upper panels) and high levels (middle and lower panels). (B) Expressions of Gli1, E-Cad and β-Catenin (β-Cat) in three representative tissue specimens in the Tianjin cohort with Gli1 expression at a low level (upper panels), a mixed expression pattern (middle panels) and a high level (lower panels). (C) Correlations between Gli1, EMT markers, and recurrence/metastasis. Statistical analysis was performed between Gli1 and E-Cad, Gli1 and β-Cat, Gli1 and recurrence/ metastasis. (D) Gli1 and E-Cad expression in four lung SCC cell lines by Western blots.
    Figure Legend Snippet: Gli1 expression reversely correlates with E-Cadherin expression in lung SCC. (A) Expressions of Gli1 and E-Cadherin (E-Cad) in three representative tissue specimens in the UCSF cohort with Gli1 expression at a low level (upper panels) and high levels (middle and lower panels). (B) Expressions of Gli1, E-Cad and β-Catenin (β-Cat) in three representative tissue specimens in the Tianjin cohort with Gli1 expression at a low level (upper panels), a mixed expression pattern (middle panels) and a high level (lower panels). (C) Correlations between Gli1, EMT markers, and recurrence/metastasis. Statistical analysis was performed between Gli1 and E-Cad, Gli1 and β-Cat, Gli1 and recurrence/ metastasis. (D) Gli1 and E-Cad expression in four lung SCC cell lines by Western blots.

    Techniques Used: Expressing, Western Blot

    Aberrant activation of the Shh pathway in lung SCC. (A) Representative protein expression of Shh, Smo, Ptch1 and Gli1 by IHC staining, scored as 0 (negative), 1 (mild positive), 2 (positive), and 3 (strong positive). (B) Expression distributions of Shh, Smo, Ptch1 and Gli1 in 177 patient tissue specimens in the Tianjin cohort. (C) Association between the expressions of Hh pathway components. Kendall’s tau-b statsitcs was used to determine the correlation between proteins. The correlation coefficient r and p values were presented in (C) . Kappa test was also performed with IHC scores of 1–3 grouped as “+”, 0 as “-”. Kappa test’s p values were labeled with*.
    Figure Legend Snippet: Aberrant activation of the Shh pathway in lung SCC. (A) Representative protein expression of Shh, Smo, Ptch1 and Gli1 by IHC staining, scored as 0 (negative), 1 (mild positive), 2 (positive), and 3 (strong positive). (B) Expression distributions of Shh, Smo, Ptch1 and Gli1 in 177 patient tissue specimens in the Tianjin cohort. (C) Association between the expressions of Hh pathway components. Kendall’s tau-b statsitcs was used to determine the correlation between proteins. The correlation coefficient r and p values were presented in (C) . Kappa test was also performed with IHC scores of 1–3 grouped as “+”, 0 as “-”. Kappa test’s p values were labeled with*.

    Techniques Used: Activation Assay, Expressing, Immunohistochemistry, Staining, Labeling

    12) Product Images from "Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *"

    Article Title: Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.606483

    Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.
    Figure Legend Snippet: Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.

    Techniques Used: Inhibition, Transfection

    Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,
    Figure Legend Snippet: Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,

    Techniques Used: Cycling Probe Technology, DNA Synthesis

    A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which
    Figure Legend Snippet: A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which

    Techniques Used: Inhibition, Cycling Probe Technology, Binding Assay, Recombinase Polymerase Amplification

    Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549
    Figure Legend Snippet: Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549

    Techniques Used: Inhibition, Cycling Probe Technology

    Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE
    Figure Legend Snippet: Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE

    Techniques Used: Inhibition, Cycling Probe Technology, Transfection, SDS Page

    Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described
    Figure Legend Snippet: Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described

    Techniques Used: Expressing, Activity Assay, In Silico, Binding Assay, Luciferase, Reporter Assay

    Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used
    Figure Legend Snippet: Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used

    Techniques Used: Cycling Probe Technology

    Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU
    Figure Legend Snippet: Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU

    Techniques Used: Inhibition, DNA Synthesis, Labeling

    13) Product Images from "Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *"

    Article Title: Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.606483

    Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.
    Figure Legend Snippet: Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.

    Techniques Used: Inhibition, Transfection

    Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,
    Figure Legend Snippet: Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,

    Techniques Used: Cycling Probe Technology, DNA Synthesis

    A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which
    Figure Legend Snippet: A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which

    Techniques Used: Inhibition, Cycling Probe Technology, Binding Assay, Recombinase Polymerase Amplification

    Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549
    Figure Legend Snippet: Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549

    Techniques Used: Inhibition, Cycling Probe Technology

    Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE
    Figure Legend Snippet: Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE

    Techniques Used: Inhibition, Cycling Probe Technology, Transfection, SDS Page

    Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described
    Figure Legend Snippet: Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described

    Techniques Used: Expressing, Activity Assay, In Silico, Binding Assay, Luciferase, Reporter Assay

    Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used
    Figure Legend Snippet: Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used

    Techniques Used: Cycling Probe Technology

    Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU
    Figure Legend Snippet: Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU

    Techniques Used: Inhibition, DNA Synthesis, Labeling

    14) Product Images from "Nonclassical Activation of Hedgehog Signaling Enhances Multidrug Resistance and Makes Cancer Cells Refractory to Smoothened-targeting Hedgehog Inhibition *"

    Article Title: Nonclassical Activation of Hedgehog Signaling Enhances Multidrug Resistance and Makes Cancer Cells Refractory to Smoothened-targeting Hedgehog Inhibition *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M112.432302

    OPN regulates GLI1 activity, expression, and induces epithelial-mesenchymal plasticity. A , OPN activates an 8×luciferase GLI reporter in MCF10AT cells (*, p
    Figure Legend Snippet: OPN regulates GLI1 activity, expression, and induces epithelial-mesenchymal plasticity. A , OPN activates an 8×luciferase GLI reporter in MCF10AT cells (*, p

    Techniques Used: Activity Assay, Expressing

    Inactivation of GSK3β by OPN causes accumulation of GLI1 in the nucleus and enhances GLI-mediated transcriptional activity. A , OPN induced phosphorylation of Akt at Ser-473 and phosphorylation of GSK3β at Ser-9. SUM1315 and T47D cells
    Figure Legend Snippet: Inactivation of GSK3β by OPN causes accumulation of GLI1 in the nucleus and enhances GLI-mediated transcriptional activity. A , OPN induced phosphorylation of Akt at Ser-473 and phosphorylation of GSK3β at Ser-9. SUM1315 and T47D cells

    Techniques Used: Activity Assay

    15) Product Images from "Microvesicles releasing by oral cancer cells enhance endothelial cell angiogenesis via Shh/RhoA signaling pathway"

    Article Title: Microvesicles releasing by oral cancer cells enhance endothelial cell angiogenesis via Shh/RhoA signaling pathway

    Journal: Cancer Biology & Therapy

    doi: 10.1080/15384047.2017.1373213

    Immunohistochemical detection of Shh, Gli-1 in metastatic lymphnodes of OSCC specimens. A, cytoplasm of expression of metastatic lymphnodes for Shh (strong staining); B, nuclear and cytoplasm expression of metastatic lymphnodes for Gli1 (strong staining). Bars indicate 100 um.
    Figure Legend Snippet: Immunohistochemical detection of Shh, Gli-1 in metastatic lymphnodes of OSCC specimens. A, cytoplasm of expression of metastatic lymphnodes for Shh (strong staining); B, nuclear and cytoplasm expression of metastatic lymphnodes for Gli1 (strong staining). Bars indicate 100 um.

    Techniques Used: Immunohistochemistry, Expressing, Staining

    Immunohistochemical detection of Shh, Gli1 in OSCC specimens. A, cytoplasm expression of OSCC specimen for Shh (a1, low staining; a2, moderate staining; a3, strong staining); B, nuclear and cytoplasm expression of OSCC specimen for Gli1 (b1, low staining; b2, moderate staining; b3, strong staining). Bars indicate 100 um.
    Figure Legend Snippet: Immunohistochemical detection of Shh, Gli1 in OSCC specimens. A, cytoplasm expression of OSCC specimen for Shh (a1, low staining; a2, moderate staining; a3, strong staining); B, nuclear and cytoplasm expression of OSCC specimen for Gli1 (b1, low staining; b2, moderate staining; b3, strong staining). Bars indicate 100 um.

    Techniques Used: Immunohistochemistry, Expressing, Staining

    The efficacy of Shh/RhoA pathway on tube formation of HUVECs in vitro . A, Phase-contrast micrographs showed that MVs harboring Shh enhanced network formation of HUVECs on Matrigel. The efficacy of MVs-enhanced network formation was inhibited by C3 transferase (5 ug/ml). The Shh (5 ug/ml) was used as the positive control. The treatment condition and the actual number of branch points ± SEM were underneath the image. Branch points were used to quantify angiogenesis; B, Shh associated tube formation was related with RhoA activation; C, the expression of Gli1 in HUVECs after rh-shh (5 ug/ml) treatment. A difference of P
    Figure Legend Snippet: The efficacy of Shh/RhoA pathway on tube formation of HUVECs in vitro . A, Phase-contrast micrographs showed that MVs harboring Shh enhanced network formation of HUVECs on Matrigel. The efficacy of MVs-enhanced network formation was inhibited by C3 transferase (5 ug/ml). The Shh (5 ug/ml) was used as the positive control. The treatment condition and the actual number of branch points ± SEM were underneath the image. Branch points were used to quantify angiogenesis; B, Shh associated tube formation was related with RhoA activation; C, the expression of Gli1 in HUVECs after rh-shh (5 ug/ml) treatment. A difference of P

    Techniques Used: In Vitro, Positive Control, Activation Assay, Expressing

    16) Product Images from "Dual degradation signals destruct GLI1: AMPK inhibits GLI1 through β-TrCP-mediated proteasome degradation"

    Article Title: Dual degradation signals destruct GLI1: AMPK inhibits GLI1 through β-TrCP-mediated proteasome degradation

    Journal: Oncotarget

    doi: 10.18632/oncotarget.17769

    GLI1 expression inversely correlates with p-AMPK in human brain cancer tissues A cohort of 198 human brain cancer samples which included 35 medulloblastoma samples tiled on a tissue array were analyzed by immunohistochemical staining with an anti-GLI1 antibody (A and C) and an anti-p-AMPK antibody (B and D) . Representative images were consecutive sections of two different MB patients. Scale bar: 50 μm
    Figure Legend Snippet: GLI1 expression inversely correlates with p-AMPK in human brain cancer tissues A cohort of 198 human brain cancer samples which included 35 medulloblastoma samples tiled on a tissue array were analyzed by immunohistochemical staining with an anti-GLI1 antibody (A and C) and an anti-p-AMPK antibody (B and D) . Representative images were consecutive sections of two different MB patients. Scale bar: 50 μm

    Techniques Used: Expressing, Immunohistochemistry, Staining

    AMPK promotes β-TrCP-mediated GLI1 ubiquitination and degradation (A) HEK293 cells were co-transfected with β-TrCP, Ub and Flag-tagged GLI1 wt for 36 hours and then treated with MG-132 (10 μM) for 6 hours prior to harvest. The lysates were immunoprecipitated and immunoblotted with anti-Flag antibody. Smear bands show GLI1 ubiquitination. (B) HEK293 cells were co-transfected with β-TrCP, Ub and Flag-tagged GLI1 wt , GLI1 3A and GLI1 3E for 36 hours and treated with MG-132 (10 μM) for 6 hours prior to harvest. Ubiquitination of GLI1 was analyzed from IP of GLI1-Flag in WB with anti-Flag antibody. (C) Flag-tagged GLI1 wt , GLI1 3A or GLI1 3E was co-transfected with Myc-tagged β-TrCP (1 and 3 μg DNA) and β-TrCP ΔF (3 μg) for 36 hours in HEK293 cells. Lysates were harvested for immunoblotting with indicated antibody.
    Figure Legend Snippet: AMPK promotes β-TrCP-mediated GLI1 ubiquitination and degradation (A) HEK293 cells were co-transfected with β-TrCP, Ub and Flag-tagged GLI1 wt for 36 hours and then treated with MG-132 (10 μM) for 6 hours prior to harvest. The lysates were immunoprecipitated and immunoblotted with anti-Flag antibody. Smear bands show GLI1 ubiquitination. (B) HEK293 cells were co-transfected with β-TrCP, Ub and Flag-tagged GLI1 wt , GLI1 3A and GLI1 3E for 36 hours and treated with MG-132 (10 μM) for 6 hours prior to harvest. Ubiquitination of GLI1 was analyzed from IP of GLI1-Flag in WB with anti-Flag antibody. (C) Flag-tagged GLI1 wt , GLI1 3A or GLI1 3E was co-transfected with Myc-tagged β-TrCP (1 and 3 μg DNA) and β-TrCP ΔF (3 μg) for 36 hours in HEK293 cells. Lysates were harvested for immunoblotting with indicated antibody.

    Techniques Used: Transfection, Immunoprecipitation, Western Blot

    GLI1 promotes MB cell growth and colony formation (A) GLI1 protein levels were determined by Western blotting in GLI1 stably overexpressed Daoy-GLI1 (A), MEF-GLI1 (B) and NIH3T3-GLI1 (C) by comparing with corresponding control cells (Inserted panel). The cell growth assay was examined by MTT assay. The line charts show the means of three independent experiments, and error bars show standard deviations. (D) The GLI1 function on anchorage- independent growth was determined by soft agar assay. The cells (DAOY-Ctrl/GLI1 and NIH3T3-Ctrl/GLI1) were seeded on agarose plates for colony formation assays, and colonies larger than 0.2 mm were counted two weeks later. The histogram shows the relative fold changes of three independent experiments, and error bars show standard deviations. (“*” p
    Figure Legend Snippet: GLI1 promotes MB cell growth and colony formation (A) GLI1 protein levels were determined by Western blotting in GLI1 stably overexpressed Daoy-GLI1 (A), MEF-GLI1 (B) and NIH3T3-GLI1 (C) by comparing with corresponding control cells (Inserted panel). The cell growth assay was examined by MTT assay. The line charts show the means of three independent experiments, and error bars show standard deviations. (D) The GLI1 function on anchorage- independent growth was determined by soft agar assay. The cells (DAOY-Ctrl/GLI1 and NIH3T3-Ctrl/GLI1) were seeded on agarose plates for colony formation assays, and colonies larger than 0.2 mm were counted two weeks later. The histogram shows the relative fold changes of three independent experiments, and error bars show standard deviations. (“*” p

    Techniques Used: Western Blot, Stable Transfection, Growth Assay, MTT Assay, Soft Agar Assay

    AMPK controls GLI1 protein localization (A) NIH3T3 GLI1-Flag stable cells were treated with Comp C (10 μM) for 6 hours and 2DG (25 mM) for 30 minutes, and cells were analyzed by immunofluorescence staining with anti-Flag antibody and DAPI. Bar graphs showed the percentages of cells with cytosolic (C) or nuclear (N) GLI1 expressions. (B) NIH3T3 GLI1-flag stable cells (GLI1 WT , GLI1 3A and GLI1 3E ) were treated with MG-132 (10 μM) for 4 hours before cells were analyzed by immunofluorescence staining with anti-Flag antibody. All the results were from three independent immunofluorescence experiments and error bars show standard deviations. (“**” p
    Figure Legend Snippet: AMPK controls GLI1 protein localization (A) NIH3T3 GLI1-Flag stable cells were treated with Comp C (10 μM) for 6 hours and 2DG (25 mM) for 30 minutes, and cells were analyzed by immunofluorescence staining with anti-Flag antibody and DAPI. Bar graphs showed the percentages of cells with cytosolic (C) or nuclear (N) GLI1 expressions. (B) NIH3T3 GLI1-flag stable cells (GLI1 WT , GLI1 3A and GLI1 3E ) were treated with MG-132 (10 μM) for 4 hours before cells were analyzed by immunofluorescence staining with anti-Flag antibody. All the results were from three independent immunofluorescence experiments and error bars show standard deviations. (“**” p

    Techniques Used: Immunofluorescence, Staining

    β-TrCP is essential for AMPK-mediated Gli1 degradation (A) Both β-TrCP wild-type ( +/+ ) and β-TrCP knockout ( −/− ) MEFs were treated with 2DG (0, 25, 50 mM) for 4 hours. Lysates were collected and analyzed by Western blot using the indicated antibody. (B) Reciprocal co-IP assays were performed with antibodies against GLI1 or β-TrCP to pull down GLI1 or β-TrCP protein in pZp53Med1 (Med1) cells. Western blots were performed with GLI1 and β-TrCP antibodies in both IP samples. (C) Flag-GLI1 was transiently transfected into HEK293 cells for 48 h and followed by treatment with 2DG (25 mM) with or without Comp C (20 μM) for 30 mins. Immunoblotting for Flag-GLI1 or β-TrCP was performed following IP of Flag-GLI1. WB from the lysate indicates that the Flag-GLI1 transfection and the compound treatments proceeded as expected as p-AMPK and p-ACC levels are increased by 2DG (lane 2) but decreased by Comp C (lane 3). The WB intensity was measured by Image J and the annotated number indicates the ratio of β-TrCP to GLI1 levels normalized to control group. (D) Immunoblot for either Flag-GLI1 or β-TrCP using lysates from the IP of β-TrCP from GLI1 wt , GLI1 3A and GLI1 3E stably expressed NIH3T3 cells with MG-132 (10 μM) for 4 hours. The WB intensity was measured by Image J and the annotated number indicates the ratio of GLI1 to β-TrCP levels normalized to Gli1 WT group.
    Figure Legend Snippet: β-TrCP is essential for AMPK-mediated Gli1 degradation (A) Both β-TrCP wild-type ( +/+ ) and β-TrCP knockout ( −/− ) MEFs were treated with 2DG (0, 25, 50 mM) for 4 hours. Lysates were collected and analyzed by Western blot using the indicated antibody. (B) Reciprocal co-IP assays were performed with antibodies against GLI1 or β-TrCP to pull down GLI1 or β-TrCP protein in pZp53Med1 (Med1) cells. Western blots were performed with GLI1 and β-TrCP antibodies in both IP samples. (C) Flag-GLI1 was transiently transfected into HEK293 cells for 48 h and followed by treatment with 2DG (25 mM) with or without Comp C (20 μM) for 30 mins. Immunoblotting for Flag-GLI1 or β-TrCP was performed following IP of Flag-GLI1. WB from the lysate indicates that the Flag-GLI1 transfection and the compound treatments proceeded as expected as p-AMPK and p-ACC levels are increased by 2DG (lane 2) but decreased by Comp C (lane 3). The WB intensity was measured by Image J and the annotated number indicates the ratio of β-TrCP to GLI1 levels normalized to control group. (D) Immunoblot for either Flag-GLI1 or β-TrCP using lysates from the IP of β-TrCP from GLI1 wt , GLI1 3A and GLI1 3E stably expressed NIH3T3 cells with MG-132 (10 μM) for 4 hours. The WB intensity was measured by Image J and the annotated number indicates the ratio of GLI1 to β-TrCP levels normalized to Gli1 WT group.

    Techniques Used: Knock-Out, Western Blot, Co-Immunoprecipitation Assay, Transfection, Stable Transfection

    17) Product Images from "Hedgehog/Gli promotes epithelial-mesenchymal transition in lung squamous cell carcinomas"

    Article Title: Hedgehog/Gli promotes epithelial-mesenchymal transition in lung squamous cell carcinomas

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/1756-9966-33-34

    Gli1 expression reversely correlates with E-Cadherin expression in lung SCC. (A) Expressions of Gli1 and E-Cadherin (E-Cad) in three representative tissue specimens in the UCSF cohort with Gli1 expression at a low level (upper panels) and high levels (middle and lower panels). (B) Expressions of Gli1, E-Cad and β-Catenin (β-Cat) in three representative tissue specimens in the Tianjin cohort with Gli1 expression at a low level (upper panels), a mixed expression pattern (middle panels) and a high level (lower panels). (C) Correlations between Gli1, EMT markers, and recurrence/metastasis. Statistical analysis was performed between Gli1 and E-Cad, Gli1 and β-Cat, Gli1 and recurrence/ metastasis. (D) Gli1 and E-Cad expression in four lung SCC cell lines by Western blots.
    Figure Legend Snippet: Gli1 expression reversely correlates with E-Cadherin expression in lung SCC. (A) Expressions of Gli1 and E-Cadherin (E-Cad) in three representative tissue specimens in the UCSF cohort with Gli1 expression at a low level (upper panels) and high levels (middle and lower panels). (B) Expressions of Gli1, E-Cad and β-Catenin (β-Cat) in three representative tissue specimens in the Tianjin cohort with Gli1 expression at a low level (upper panels), a mixed expression pattern (middle panels) and a high level (lower panels). (C) Correlations between Gli1, EMT markers, and recurrence/metastasis. Statistical analysis was performed between Gli1 and E-Cad, Gli1 and β-Cat, Gli1 and recurrence/ metastasis. (D) Gli1 and E-Cad expression in four lung SCC cell lines by Western blots.

    Techniques Used: Expressing, Western Blot

    Aberrant activation of the Shh pathway in lung SCC. (A) Representative protein expression of Shh, Smo, Ptch1 and Gli1 by IHC staining, scored as 0 (negative), 1 (mild positive), 2 (positive), and 3 (strong positive). (B) Expression distributions of Shh, Smo, Ptch1 and Gli1 in 177 patient tissue specimens in the Tianjin cohort. (C) Association between the expressions of Hh pathway components. Kendall’s tau-b statsitcs was used to determine the correlation between proteins. The correlation coefficient r and p values were presented in (C) . Kappa test was also performed with IHC scores of 1–3 grouped as “+”, 0 as “-”. Kappa test’s p values were labeled with*.
    Figure Legend Snippet: Aberrant activation of the Shh pathway in lung SCC. (A) Representative protein expression of Shh, Smo, Ptch1 and Gli1 by IHC staining, scored as 0 (negative), 1 (mild positive), 2 (positive), and 3 (strong positive). (B) Expression distributions of Shh, Smo, Ptch1 and Gli1 in 177 patient tissue specimens in the Tianjin cohort. (C) Association between the expressions of Hh pathway components. Kendall’s tau-b statsitcs was used to determine the correlation between proteins. The correlation coefficient r and p values were presented in (C) . Kappa test was also performed with IHC scores of 1–3 grouped as “+”, 0 as “-”. Kappa test’s p values were labeled with*.

    Techniques Used: Activation Assay, Expressing, Immunohistochemistry, Staining, Labeling

    18) Product Images from "Gli2 Acetylation at Lysine 757 Regulates Hedgehog-Dependent Transcriptional Output by Preventing Its Promoter Occupancy"

    Article Title: Gli2 Acetylation at Lysine 757 Regulates Hedgehog-Dependent Transcriptional Output by Preventing Its Promoter Occupancy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0065718

    Model of acetylation-dependent control of Gli activity. Following Hh/Ptch interaction, Smo triggers a signaling cascade leading to Gli2 deacetylation and to the inhibition of the βTrCP-regulated balance between Gli2R and full length Gli2 (Gli2A). Both events contributes to the early signal-dependent activation of the Hh pathway. Once activated, Gli2 promotes transcription of Gli1, whose activity is also regulated by Hh-induced HDAC1-mediated deacetylation, thus generating a positive feedback loop (late activation).
    Figure Legend Snippet: Model of acetylation-dependent control of Gli activity. Following Hh/Ptch interaction, Smo triggers a signaling cascade leading to Gli2 deacetylation and to the inhibition of the βTrCP-regulated balance between Gli2R and full length Gli2 (Gli2A). Both events contributes to the early signal-dependent activation of the Hh pathway. Once activated, Gli2 promotes transcription of Gli1, whose activity is also regulated by Hh-induced HDAC1-mediated deacetylation, thus generating a positive feedback loop (late activation).

    Techniques Used: Activity Assay, Inhibition, Activation Assay

    19) Product Images from "Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *"

    Article Title: Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.606483

    Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.
    Figure Legend Snippet: Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.

    Techniques Used: Inhibition, Transfection

    Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,
    Figure Legend Snippet: Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,

    Techniques Used: Cycling Probe Technology, DNA Synthesis

    A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which
    Figure Legend Snippet: A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which

    Techniques Used: Inhibition, Cycling Probe Technology, Binding Assay, Recombinase Polymerase Amplification

    Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549
    Figure Legend Snippet: Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549

    Techniques Used: Inhibition, Cycling Probe Technology

    Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE
    Figure Legend Snippet: Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE

    Techniques Used: Inhibition, Cycling Probe Technology, Transfection, SDS Page

    Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described
    Figure Legend Snippet: Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described

    Techniques Used: Expressing, Activity Assay, In Silico, Binding Assay, Luciferase, Reporter Assay

    Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used
    Figure Legend Snippet: Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used

    Techniques Used: Cycling Probe Technology

    Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU
    Figure Legend Snippet: Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU

    Techniques Used: Inhibition, DNA Synthesis, Labeling

    20) Product Images from "Microvesicles releasing by oral cancer cells enhance endothelial cell angiogenesis via Shh/RhoA signaling pathway"

    Article Title: Microvesicles releasing by oral cancer cells enhance endothelial cell angiogenesis via Shh/RhoA signaling pathway

    Journal: Cancer Biology & Therapy

    doi: 10.1080/15384047.2017.1373213

    Immunohistochemical detection of Shh, Gli-1 in metastatic lymphnodes of OSCC specimens. A, cytoplasm of expression of metastatic lymphnodes for Shh (strong staining); B, nuclear and cytoplasm expression of metastatic lymphnodes for Gli1 (strong staining). Bars indicate 100 um.
    Figure Legend Snippet: Immunohistochemical detection of Shh, Gli-1 in metastatic lymphnodes of OSCC specimens. A, cytoplasm of expression of metastatic lymphnodes for Shh (strong staining); B, nuclear and cytoplasm expression of metastatic lymphnodes for Gli1 (strong staining). Bars indicate 100 um.

    Techniques Used: Immunohistochemistry, Expressing, Staining

    Immunohistochemical detection of Shh, Gli1 in OSCC specimens. A, cytoplasm expression of OSCC specimen for Shh (a1, low staining; a2, moderate staining; a3, strong staining); B, nuclear and cytoplasm expression of OSCC specimen for Gli1 (b1, low staining; b2, moderate staining; b3, strong staining). Bars indicate 100 um.
    Figure Legend Snippet: Immunohistochemical detection of Shh, Gli1 in OSCC specimens. A, cytoplasm expression of OSCC specimen for Shh (a1, low staining; a2, moderate staining; a3, strong staining); B, nuclear and cytoplasm expression of OSCC specimen for Gli1 (b1, low staining; b2, moderate staining; b3, strong staining). Bars indicate 100 um.

    Techniques Used: Immunohistochemistry, Expressing, Staining

    The efficacy of Shh/RhoA pathway on tube formation of HUVECs in vitro . A, Phase-contrast micrographs showed that MVs harboring Shh enhanced network formation of HUVECs on Matrigel. The efficacy of MVs-enhanced network formation was inhibited by C3 transferase (5 ug/ml). The Shh (5 ug/ml) was used as the positive control. The treatment condition and the actual number of branch points ± SEM were underneath the image. Branch points were used to quantify angiogenesis; B, Shh associated tube formation was related with RhoA activation; C, the expression of Gli1 in HUVECs after rh-shh (5 ug/ml) treatment. A difference of P
    Figure Legend Snippet: The efficacy of Shh/RhoA pathway on tube formation of HUVECs in vitro . A, Phase-contrast micrographs showed that MVs harboring Shh enhanced network formation of HUVECs on Matrigel. The efficacy of MVs-enhanced network formation was inhibited by C3 transferase (5 ug/ml). The Shh (5 ug/ml) was used as the positive control. The treatment condition and the actual number of branch points ± SEM were underneath the image. Branch points were used to quantify angiogenesis; B, Shh associated tube formation was related with RhoA activation; C, the expression of Gli1 in HUVECs after rh-shh (5 ug/ml) treatment. A difference of P

    Techniques Used: In Vitro, Positive Control, Activation Assay, Expressing

    21) Product Images from "Trimeric G protein-CARMA1 axis links smoothened, the hedgehog receptor transducer, to NF-?B activation in diffuse large B-cell lymphoma"

    Article Title: Trimeric G protein-CARMA1 axis links smoothened, the hedgehog receptor transducer, to NF-?B activation in diffuse large B-cell lymphoma

    Journal: Blood

    doi: 10.1182/blood-2012-12-470153

    SMO activates NF-κB pathway independent of GLI1. (A) Silencing SMO in DOHH2 and HBL1 resulted in decrease of p -p65, (B) decrease of nuclear DNA binding activity of p65 and p50, and (C) decrease of total NF-κB luciferase activity. (D) Silencing
    Figure Legend Snippet: SMO activates NF-κB pathway independent of GLI1. (A) Silencing SMO in DOHH2 and HBL1 resulted in decrease of p -p65, (B) decrease of nuclear DNA binding activity of p65 and p50, and (C) decrease of total NF-κB luciferase activity. (D) Silencing

    Techniques Used: Binding Assay, Activity Assay, Luciferase

    Hh and NF-κB pathways are positively correlated in DLBCL, and Hh pathway contributes to NF-κB activation in DLBCL. (A) GLI1 expression levels in 145 DLBCL tumor biopsies. (B) High GLI1 expression was more frequent in DLBCL of ABC type
    Figure Legend Snippet: Hh and NF-κB pathways are positively correlated in DLBCL, and Hh pathway contributes to NF-κB activation in DLBCL. (A) GLI1 expression levels in 145 DLBCL tumor biopsies. (B) High GLI1 expression was more frequent in DLBCL of ABC type

    Techniques Used: Activation Assay, Expressing

    22) Product Images from "The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *"

    Article Title: The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M109.021949

    The Hh pathway transcriptionally regulates OPN. A , levels of GLI1 and OPN are increased in primary cutaneous cancer and metastatic melanoma. Gene microarray analysis (utilizing a Human Genome U133 Plus 2.0 array from Affymetrix, Inc.) was used to compare
    Figure Legend Snippet: The Hh pathway transcriptionally regulates OPN. A , levels of GLI1 and OPN are increased in primary cutaneous cancer and metastatic melanoma. Gene microarray analysis (utilizing a Human Genome U133 Plus 2.0 array from Affymetrix, Inc.) was used to compare

    Techniques Used: Microarray

    Silencing endogenous GLI1 expression diminishes attributes of motility, invasion, migration, and proliferation and negatively impacts tumorigenicity and metastasis. A , abrogating GLI1 expression ( KO1 and KO4 ) reduced the ability of cells to move and fill
    Figure Legend Snippet: Silencing endogenous GLI1 expression diminishes attributes of motility, invasion, migration, and proliferation and negatively impacts tumorigenicity and metastasis. A , abrogating GLI1 expression ( KO1 and KO4 ) reduced the ability of cells to move and fill

    Techniques Used: Expressing, Migration

    The OPN promoter bears a GLI1-binding site that responds to GLI1. A , mutating the putative GLI1-binding site in the OPN promoter makes the OPN promoter insensitive to the effects of GLI1. The OPN promoter (+112 to −352 (pGL3-OPN-352)) was significantly
    Figure Legend Snippet: The OPN promoter bears a GLI1-binding site that responds to GLI1. A , mutating the putative GLI1-binding site in the OPN promoter makes the OPN promoter insensitive to the effects of GLI1. The OPN promoter (+112 to −352 (pGL3-OPN-352)) was significantly

    Techniques Used: Binding Assay

    Restoring the availability of OPN-initiated signaling in GLI1-silenced cells reinstates their motility and ability to migrate and invade. A , immunoblot representing the restored OPN in cells that have been stably knocked down for GLI1 (GLI1 KO; and consequently
    Figure Legend Snippet: Restoring the availability of OPN-initiated signaling in GLI1-silenced cells reinstates their motility and ability to migrate and invade. A , immunoblot representing the restored OPN in cells that have been stably knocked down for GLI1 (GLI1 KO; and consequently

    Techniques Used: Stable Transfection

    shRNA to GLI1 abrogates expression of OPN and brings about a partial reversal of EMT. A , MDA-MB-435 cells were stably transfected with shRNA to GLI1. Clones KO1 to KO4 were stably silenced for GLI1 and show notably reduced OPN expression. B , real time
    Figure Legend Snippet: shRNA to GLI1 abrogates expression of OPN and brings about a partial reversal of EMT. A , MDA-MB-435 cells were stably transfected with shRNA to GLI1. Clones KO1 to KO4 were stably silenced for GLI1 and show notably reduced OPN expression. B , real time

    Techniques Used: shRNA, Expressing, Multiple Displacement Amplification, Stable Transfection, Transfection, Clone Assay

    23) Product Images from "Defining Causative Factors Contributing in the Activation of Hedgehog Signaling in Diffuse Large B-Cell Lymphoma"

    Article Title: Defining Causative Factors Contributing in the Activation of Hedgehog Signaling in Diffuse Large B-Cell Lymphoma

    Journal: Leukemia research

    doi: 10.1016/j.leukres.2012.06.014

    The activation status of PI3K and NF-kB modulates GLI1 and Hh levels in DLBCL cell lines
    Figure Legend Snippet: The activation status of PI3K and NF-kB modulates GLI1 and Hh levels in DLBCL cell lines

    Techniques Used: Activation Assay

    24) Product Images from "LARP7 in papillary thyroid carcinoma induces NIS expression through suppression of the SHH signaling pathway"

    Article Title: LARP7 in papillary thyroid carcinoma induces NIS expression through suppression of the SHH signaling pathway

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2018.8856

    LARP7 inhibited the activation of the SHH signaling pathway. TPC-1 and BCPAP cells were transfected with Lv-control or Lv-LARP7, and the mRNA expression levels of (A) SHH, (B) GLI1 and (C) PTCH1 and their (D) protein expression levels were quantified using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the mean ± standard deviation. *P
    Figure Legend Snippet: LARP7 inhibited the activation of the SHH signaling pathway. TPC-1 and BCPAP cells were transfected with Lv-control or Lv-LARP7, and the mRNA expression levels of (A) SHH, (B) GLI1 and (C) PTCH1 and their (D) protein expression levels were quantified using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the mean ± standard deviation. *P

    Techniques Used: Activation Assay, Transfection, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation

    25) Product Images from "Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *"

    Article Title: Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.606483

    Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.
    Figure Legend Snippet: Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.

    Techniques Used: Inhibition, Transfection

    Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,
    Figure Legend Snippet: Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,

    Techniques Used: Cycling Probe Technology, DNA Synthesis

    A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which
    Figure Legend Snippet: A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which

    Techniques Used: Inhibition, Cycling Probe Technology, Binding Assay, Recombinase Polymerase Amplification

    Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549
    Figure Legend Snippet: Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549

    Techniques Used: Inhibition, Cycling Probe Technology

    Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE
    Figure Legend Snippet: Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE

    Techniques Used: Inhibition, Cycling Probe Technology, Transfection, SDS Page

    Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described
    Figure Legend Snippet: Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described

    Techniques Used: Expressing, Activity Assay, In Silico, Binding Assay, Luciferase, Reporter Assay

    Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used
    Figure Legend Snippet: Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used

    Techniques Used: Cycling Probe Technology

    Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU
    Figure Legend Snippet: Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU

    Techniques Used: Inhibition, DNA Synthesis, Labeling

    26) Product Images from "Rhein, a Natural Anthraquinone Derivative, Attenuates the Activation of Pancreatic Stellate Cells and Ameliorates Pancreatic Fibrosis in Mice with Experimental Chronic Pancreatitis"

    Article Title: Rhein, a Natural Anthraquinone Derivative, Attenuates the Activation of Pancreatic Stellate Cells and Ameliorates Pancreatic Fibrosis in Mice with Experimental Chronic Pancreatitis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0082201

    Rhein inhibits SHH and NF-κB signals in-vitro . (A) LTC-14 cells were treated with recombinant TGF-β (0, 1, 5 or 10 ng/mL) or SHH (0, 10, 50 or 100 ng/mL) in SFM for 24 hours and subjected to Western blotting analyses. Immunoblots were probed with anti-SHH antibody for the TGF-β treatment or probed with anti-α-SMA antibody for the SHH treatment. β-ACTIN was served as a loading reference. (B) LTC-14 cells were treated with TGF-β (0, 1, 5 or 10 ng/mL) in SFM for 24 hours and harvested for mRNA extraction. Transcripts of Gli1 and Gli2 were amplified by means of qPCR, normalized to the endogenous reference Gapdh and expressed as fold changes over the non-TGF-β-treated control (SFM control). * p
    Figure Legend Snippet: Rhein inhibits SHH and NF-κB signals in-vitro . (A) LTC-14 cells were treated with recombinant TGF-β (0, 1, 5 or 10 ng/mL) or SHH (0, 10, 50 or 100 ng/mL) in SFM for 24 hours and subjected to Western blotting analyses. Immunoblots were probed with anti-SHH antibody for the TGF-β treatment or probed with anti-α-SMA antibody for the SHH treatment. β-ACTIN was served as a loading reference. (B) LTC-14 cells were treated with TGF-β (0, 1, 5 or 10 ng/mL) in SFM for 24 hours and harvested for mRNA extraction. Transcripts of Gli1 and Gli2 were amplified by means of qPCR, normalized to the endogenous reference Gapdh and expressed as fold changes over the non-TGF-β-treated control (SFM control). * p

    Techniques Used: In Vitro, Recombinant, Western Blot, Amplification, Real-time Polymerase Chain Reaction

    A schematic flow chart showing the roles of TGF-β and SHH/GLI1 signaling pathways in pancreatic fibrosis.
    Figure Legend Snippet: A schematic flow chart showing the roles of TGF-β and SHH/GLI1 signaling pathways in pancreatic fibrosis.

    Techniques Used: Flow Cytometry

    Rhein suppresses SHH signaling in-vivo. (A) The fluorescent images demonstrated the immunoreactivites of GLI1 (green; FITC) and SHH (red; Rhodamine) in paraffin-embedded pancreatic tissues in which nuclei were stained blue with DAPI. (B) Protein levels of GLI1 and SHH in pancreatic homogenates of the mice were visualized on immunoblots probed with anti-GLI1 and anti-SHH antibodies whereas β-ACTIN was served as the loading reference. (C) Integrated densities of the immunobands of SHH and GLI1 were measured. Readings were normalized to the internal reference β-ACTIN and expressed as fold change over control. (D) Transcripts of Gli1 and Gli2 were amplified by means of qPCR, normalized to the endogenous reference Gapdh and expressed as fold changes over the Control group. * p
    Figure Legend Snippet: Rhein suppresses SHH signaling in-vivo. (A) The fluorescent images demonstrated the immunoreactivites of GLI1 (green; FITC) and SHH (red; Rhodamine) in paraffin-embedded pancreatic tissues in which nuclei were stained blue with DAPI. (B) Protein levels of GLI1 and SHH in pancreatic homogenates of the mice were visualized on immunoblots probed with anti-GLI1 and anti-SHH antibodies whereas β-ACTIN was served as the loading reference. (C) Integrated densities of the immunobands of SHH and GLI1 were measured. Readings were normalized to the internal reference β-ACTIN and expressed as fold change over control. (D) Transcripts of Gli1 and Gli2 were amplified by means of qPCR, normalized to the endogenous reference Gapdh and expressed as fold changes over the Control group. * p

    Techniques Used: In Vivo, Staining, Mouse Assay, Western Blot, Amplification, Real-time Polymerase Chain Reaction

    27) Product Images from "Trimeric G protein-CARMA1 axis links smoothened, the hedgehog receptor transducer, to NF-?B activation in diffuse large B-cell lymphoma"

    Article Title: Trimeric G protein-CARMA1 axis links smoothened, the hedgehog receptor transducer, to NF-?B activation in diffuse large B-cell lymphoma

    Journal: Blood

    doi: 10.1182/blood-2012-12-470153

    SMO activates NF-κB pathway independent of GLI1. (A) Silencing SMO in DOHH2 and HBL1 resulted in decrease of p -p65, (B) decrease of nuclear DNA binding activity of p65 and p50, and (C) decrease of total NF-κB luciferase activity. (D) Silencing
    Figure Legend Snippet: SMO activates NF-κB pathway independent of GLI1. (A) Silencing SMO in DOHH2 and HBL1 resulted in decrease of p -p65, (B) decrease of nuclear DNA binding activity of p65 and p50, and (C) decrease of total NF-κB luciferase activity. (D) Silencing

    Techniques Used: Binding Assay, Activity Assay, Luciferase

    Hh and NF-κB pathways are positively correlated in DLBCL, and Hh pathway contributes to NF-κB activation in DLBCL. (A) GLI1 expression levels in 145 DLBCL tumor biopsies. (B) High GLI1 expression was more frequent in DLBCL of ABC type
    Figure Legend Snippet: Hh and NF-κB pathways are positively correlated in DLBCL, and Hh pathway contributes to NF-κB activation in DLBCL. (A) GLI1 expression levels in 145 DLBCL tumor biopsies. (B) High GLI1 expression was more frequent in DLBCL of ABC type

    Techniques Used: Activation Assay, Expressing

    28) Product Images from "Capsaicin Treatment Attenuates Cholangiocarcinoma Carcinogenesis"

    Article Title: Capsaicin Treatment Attenuates Cholangiocarcinoma Carcinogenesis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0095605

    Capsaicin targets Hedgehog signaling. The expression levels of Smo, Gli1 and Gli2 were analyzed by semiquantitative RT-PCR both in SZ-1 (A) and in TFK-1 cells (B) after treatment either with control (DMSO) or capsaicin (150 µM, 200 µM) for 24 h, 48 h and 96 h. A reduction of transmembrane protein Smo was seen in both cell lines after 96 h. Capsaicin down-regulates Hedgehog targets Gli1 and Gli2 in a time-dependent manner (96 h).
    Figure Legend Snippet: Capsaicin targets Hedgehog signaling. The expression levels of Smo, Gli1 and Gli2 were analyzed by semiquantitative RT-PCR both in SZ-1 (A) and in TFK-1 cells (B) after treatment either with control (DMSO) or capsaicin (150 µM, 200 µM) for 24 h, 48 h and 96 h. A reduction of transmembrane protein Smo was seen in both cell lines after 96 h. Capsaicin down-regulates Hedgehog targets Gli1 and Gli2 in a time-dependent manner (96 h).

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    29) Product Images from "The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *"

    Article Title: The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M109.021949

    The Hh pathway transcriptionally regulates OPN. A , levels of GLI1 and OPN are increased in primary cutaneous cancer and metastatic melanoma. Gene microarray analysis (utilizing a Human Genome U133 Plus 2.0 array from Affymetrix, Inc.) was used to compare
    Figure Legend Snippet: The Hh pathway transcriptionally regulates OPN. A , levels of GLI1 and OPN are increased in primary cutaneous cancer and metastatic melanoma. Gene microarray analysis (utilizing a Human Genome U133 Plus 2.0 array from Affymetrix, Inc.) was used to compare

    Techniques Used: Microarray

    Silencing endogenous GLI1 expression diminishes attributes of motility, invasion, migration, and proliferation and negatively impacts tumorigenicity and metastasis. A , abrogating GLI1 expression ( KO1 and KO4 ) reduced the ability of cells to move and fill
    Figure Legend Snippet: Silencing endogenous GLI1 expression diminishes attributes of motility, invasion, migration, and proliferation and negatively impacts tumorigenicity and metastasis. A , abrogating GLI1 expression ( KO1 and KO4 ) reduced the ability of cells to move and fill

    Techniques Used: Expressing, Migration

    The OPN promoter bears a GLI1-binding site that responds to GLI1. A , mutating the putative GLI1-binding site in the OPN promoter makes the OPN promoter insensitive to the effects of GLI1. The OPN promoter (+112 to −352 (pGL3-OPN-352)) was significantly
    Figure Legend Snippet: The OPN promoter bears a GLI1-binding site that responds to GLI1. A , mutating the putative GLI1-binding site in the OPN promoter makes the OPN promoter insensitive to the effects of GLI1. The OPN promoter (+112 to −352 (pGL3-OPN-352)) was significantly

    Techniques Used: Binding Assay

    Restoring the availability of OPN-initiated signaling in GLI1-silenced cells reinstates their motility and ability to migrate and invade. A , immunoblot representing the restored OPN in cells that have been stably knocked down for GLI1 (GLI1 KO; and consequently
    Figure Legend Snippet: Restoring the availability of OPN-initiated signaling in GLI1-silenced cells reinstates their motility and ability to migrate and invade. A , immunoblot representing the restored OPN in cells that have been stably knocked down for GLI1 (GLI1 KO; and consequently

    Techniques Used: Stable Transfection

    shRNA to GLI1 abrogates expression of OPN and brings about a partial reversal of EMT. A , MDA-MB-435 cells were stably transfected with shRNA to GLI1. Clones KO1 to KO4 were stably silenced for GLI1 and show notably reduced OPN expression. B , real time
    Figure Legend Snippet: shRNA to GLI1 abrogates expression of OPN and brings about a partial reversal of EMT. A , MDA-MB-435 cells were stably transfected with shRNA to GLI1. Clones KO1 to KO4 were stably silenced for GLI1 and show notably reduced OPN expression. B , real time

    Techniques Used: shRNA, Expressing, Multiple Displacement Amplification, Stable Transfection, Transfection, Clone Assay

    30) Product Images from "Defining Causative Factors Contributing in the Activation of Hedgehog Signaling in Diffuse Large B-Cell Lymphoma"

    Article Title: Defining Causative Factors Contributing in the Activation of Hedgehog Signaling in Diffuse Large B-Cell Lymphoma

    Journal: Leukemia research

    doi: 10.1016/j.leukres.2012.06.014

    The activation status of PI3K and NF-kB modulates GLI1 and Hh levels in DLBCL cell lines
    Figure Legend Snippet: The activation status of PI3K and NF-kB modulates GLI1 and Hh levels in DLBCL cell lines

    Techniques Used: Activation Assay

    31) Product Images from "Defining Causative Factors Contributing in the Activation of Hedgehog Signaling in Diffuse Large B-Cell Lymphoma"

    Article Title: Defining Causative Factors Contributing in the Activation of Hedgehog Signaling in Diffuse Large B-Cell Lymphoma

    Journal: Leukemia research

    doi: 10.1016/j.leukres.2012.06.014

    The activation status of PI3K and NF-kB modulates GLI1 and Hh levels in DLBCL cell lines
    Figure Legend Snippet: The activation status of PI3K and NF-kB modulates GLI1 and Hh levels in DLBCL cell lines

    Techniques Used: Activation Assay

    32) Product Images from "Hedgehog/Gli promotes epithelial-mesenchymal transition in lung squamous cell carcinomas"

    Article Title: Hedgehog/Gli promotes epithelial-mesenchymal transition in lung squamous cell carcinomas

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/1756-9966-33-34

    Gli1 expression reversely correlates with E-Cadherin expression in lung SCC. (A) Expressions of Gli1 and E-Cadherin (E-Cad) in three representative tissue specimens in the UCSF cohort with Gli1 expression at a low level (upper panels) and high levels (middle and lower panels). (B) Expressions of Gli1, E-Cad and β-Catenin (β-Cat) in three representative tissue specimens in the Tianjin cohort with Gli1 expression at a low level (upper panels), a mixed expression pattern (middle panels) and a high level (lower panels). (C) Correlations between Gli1, EMT markers, and recurrence/metastasis. Statistical analysis was performed between Gli1 and E-Cad, Gli1 and β-Cat, Gli1 and recurrence/ metastasis. (D) Gli1 and E-Cad expression in four lung SCC cell lines by Western blots.
    Figure Legend Snippet: Gli1 expression reversely correlates with E-Cadherin expression in lung SCC. (A) Expressions of Gli1 and E-Cadherin (E-Cad) in three representative tissue specimens in the UCSF cohort with Gli1 expression at a low level (upper panels) and high levels (middle and lower panels). (B) Expressions of Gli1, E-Cad and β-Catenin (β-Cat) in three representative tissue specimens in the Tianjin cohort with Gli1 expression at a low level (upper panels), a mixed expression pattern (middle panels) and a high level (lower panels). (C) Correlations between Gli1, EMT markers, and recurrence/metastasis. Statistical analysis was performed between Gli1 and E-Cad, Gli1 and β-Cat, Gli1 and recurrence/ metastasis. (D) Gli1 and E-Cad expression in four lung SCC cell lines by Western blots.

    Techniques Used: Expressing, Western Blot

    Aberrant activation of the Shh pathway in lung SCC. (A) Representative protein expression of Shh, Smo, Ptch1 and Gli1 by IHC staining, scored as 0 (negative), 1 (mild positive), 2 (positive), and 3 (strong positive). (B) Expression distributions of Shh, Smo, Ptch1 and Gli1 in 177 patient tissue specimens in the Tianjin cohort. (C) Association between the expressions of Hh pathway components. Kendall’s tau-b statsitcs was used to determine the correlation between proteins. The correlation coefficient r and p values were presented in (C) . Kappa test was also performed with IHC scores of 1–3 grouped as “+”, 0 as “-”. Kappa test’s p values were labeled with*.
    Figure Legend Snippet: Aberrant activation of the Shh pathway in lung SCC. (A) Representative protein expression of Shh, Smo, Ptch1 and Gli1 by IHC staining, scored as 0 (negative), 1 (mild positive), 2 (positive), and 3 (strong positive). (B) Expression distributions of Shh, Smo, Ptch1 and Gli1 in 177 patient tissue specimens in the Tianjin cohort. (C) Association between the expressions of Hh pathway components. Kendall’s tau-b statsitcs was used to determine the correlation between proteins. The correlation coefficient r and p values were presented in (C) . Kappa test was also performed with IHC scores of 1–3 grouped as “+”, 0 as “-”. Kappa test’s p values were labeled with*.

    Techniques Used: Activation Assay, Expressing, Immunohistochemistry, Staining, Labeling

    33) Product Images from "Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *"

    Article Title: Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.606483

    Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.
    Figure Legend Snippet: Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.

    Techniques Used: Inhibition, Transfection

    Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,
    Figure Legend Snippet: Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,

    Techniques Used: Cycling Probe Technology, DNA Synthesis

    A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which
    Figure Legend Snippet: A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which

    Techniques Used: Inhibition, Cycling Probe Technology, Binding Assay, Recombinase Polymerase Amplification

    Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549
    Figure Legend Snippet: Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549

    Techniques Used: Inhibition, Cycling Probe Technology

    Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE
    Figure Legend Snippet: Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE

    Techniques Used: Inhibition, Cycling Probe Technology, Transfection, SDS Page

    Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described
    Figure Legend Snippet: Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described

    Techniques Used: Expressing, Activity Assay, In Silico, Binding Assay, Luciferase, Reporter Assay

    Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used
    Figure Legend Snippet: Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used

    Techniques Used: Cycling Probe Technology

    Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU
    Figure Legend Snippet: Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU

    Techniques Used: Inhibition, DNA Synthesis, Labeling

    34) Product Images from "Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *"

    Article Title: Gli1 Protein Regulates the S-phase Checkpoint in Tumor Cells via Bid Protein, and Its Inhibition Sensitizes to DNA Topoisomerase 1 Inhibitors *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.606483

    Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.
    Figure Legend Snippet: Inhibition of Gli1 in cancer cells induces DDR and attenuates their clonogenic potential. A , A549 cells were transfected with control ( siCon ) or Gli1-specific ( siGli1 ) siRNAs, and, after 48 h post-transfection, cells were analyzed for γH2AX foci.

    Techniques Used: Inhibition, Transfection

    Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,
    Figure Legend Snippet: Gli1 down-regulation inhibits the S-phase checkpoint, induces CPT-resistant DNA synthesis, and increases the number of replication-coupled DSBs. A , control and Gli1 down-regulated A549 cells were treated with the indicated concentrations of CPT or DMSO,

    Techniques Used: Cycling Probe Technology, DNA Synthesis

    A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which
    Figure Legend Snippet: A putative model showing the mechanism of Gli1 inhibition-mediated S-phase checkpoint abrogation. In response to CPT-induced replication stress or stalled replication forks, Bid binding to RPA facilitates its interaction with the ATRIP-ATR complex, which

    Techniques Used: Inhibition, Cycling Probe Technology, Binding Assay, Recombinase Polymerase Amplification

    Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549
    Figure Legend Snippet: Gli1 inhibition affects the S-phase checkpoint by down-regulation of Bid. A and B , Gli1-proficient and knockdown tumor cells were exposed to DMSO or 500 n m CPT and assessed for different DDR proteins involved in the ATR/Chk1 signaling cascade in A549

    Techniques Used: Inhibition, Cycling Probe Technology

    Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE
    Figure Legend Snippet: Gli1 inhibition abrogates Chk1 phosphorylation and sensitizes cancer cells to CPT. A and B , after 48 h of siRNA transfection, cells were treated with DMSO or 500 n m CPT for 2 h. Cell lysates were normalized for protein concentrations and analyzed by SDS-PAGE

    Techniques Used: Inhibition, Cycling Probe Technology, Transfection, SDS Page

    Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described
    Figure Legend Snippet: Gli1 regulates Bid expression in tumor cells through its promoter activity. A , in silico analysis of the BID promoter region (5′ UTR) identified a consensus Gli1 binding site ( boldface ). B , luciferase reporter assay were performed as described

    Techniques Used: Expressing, Activity Assay, In Silico, Binding Assay, Luciferase, Reporter Assay

    Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used
    Figure Legend Snippet: Gli1 knockdown affects the interaction of RPA70 with ATR-ATRIP protein complexes and attenuates the CPT-induced S-phase checkpoint response via Bid. A , control and Gli1 down-regulated A549 cells were exposed to CPT or DMSO. The protein lysates were used

    Techniques Used: Cycling Probe Technology

    Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU
    Figure Legend Snippet: Gli1 inhibition-mediated abrogation of Chk1 phosphorylation is not due to the defects in DNA synthesis. A , control and Gli1 down-regulated A549 cells were treated with 10 μ m BrdU for 30 min, and cells were labeled with FITC-conjugated anti-BrdU

    Techniques Used: Inhibition, DNA Synthesis, Labeling

    35) Product Images from "Shh and p50/Bcl3 signaling crosstalk drives pathogenesis of BCCs in gorlin syndrome"

    Article Title: Shh and p50/Bcl3 signaling crosstalk drives pathogenesis of BCCs in gorlin syndrome

    Journal: Oncotarget

    doi:

    Crosstalk of Shh and Bcl3 signaling pathways in BCC cells regulate proliferation A. mRNA expression of N-cadherin and cyclin D1 in skin of UVB-irradiated and Shh-inhibitors-treated. B. Percentage of ASZ001 cells showing nuclear localization of Bcl3 in GANT61 (10 μm) and Cyclopamine (5 μm)-treated cells. C. TaqMan real-time PCR showing effects of Isopropyl β-D-1-thiogalactopyranoside (IPTG) treatment on the expression of Bcl3, CyclinD1 and N-cadherin in ASZ001 cells. ASZ001 cells have been infected with IPTG-inducible Bcl3 shRNA lentiviral vector and were selected following treatment with puromycin containing growth medium. D. TaqMan real-time PCR showing the expression of shh signaling related genes Gli1, Gli2 and Ptch1 following knockdown of Bcl3 in ASZ001 cells. E. Colony formation assay in control, IPTG (1 mM), Cyclopamine (5 μm) and IPTG+Cyclopamine treated ASZ001 cells. F. Bar diagram showing colony numbers following various treatments.
    Figure Legend Snippet: Crosstalk of Shh and Bcl3 signaling pathways in BCC cells regulate proliferation A. mRNA expression of N-cadherin and cyclin D1 in skin of UVB-irradiated and Shh-inhibitors-treated. B. Percentage of ASZ001 cells showing nuclear localization of Bcl3 in GANT61 (10 μm) and Cyclopamine (5 μm)-treated cells. C. TaqMan real-time PCR showing effects of Isopropyl β-D-1-thiogalactopyranoside (IPTG) treatment on the expression of Bcl3, CyclinD1 and N-cadherin in ASZ001 cells. ASZ001 cells have been infected with IPTG-inducible Bcl3 shRNA lentiviral vector and were selected following treatment with puromycin containing growth medium. D. TaqMan real-time PCR showing the expression of shh signaling related genes Gli1, Gli2 and Ptch1 following knockdown of Bcl3 in ASZ001 cells. E. Colony formation assay in control, IPTG (1 mM), Cyclopamine (5 μm) and IPTG+Cyclopamine treated ASZ001 cells. F. Bar diagram showing colony numbers following various treatments.

    Techniques Used: Expressing, Irradiation, Real-time Polymerase Chain Reaction, Infection, shRNA, Plasmid Preparation, Colony Assay

    36) Product Images from "Sonic hedgehog regulation of cavernous nerve regeneration and neurite formation in aged pelvic plexus"

    Article Title: Sonic hedgehog regulation of cavernous nerve regeneration and neurite formation in aged pelvic plexus

    Journal: Experimental neurology

    doi: 10.1016/j.expneurol.2018.11.001

    IHC analysis was performed on uninjured adult and aged rat MPG/CN that was grown in organ culture for 3 days and were assayed for GLI1–3. All three GLI isoforms were identified in glial cells surrounding MPG neurons of adult and aged rats. Arrows indicate GLI1–3 proteins. 1000X magnification.
    Figure Legend Snippet: IHC analysis was performed on uninjured adult and aged rat MPG/CN that was grown in organ culture for 3 days and were assayed for GLI1–3. All three GLI isoforms were identified in glial cells surrounding MPG neurons of adult and aged rats. Arrows indicate GLI1–3 proteins. 1000X magnification.

    Techniques Used: Immunohistochemistry, Organ Culture

    37) Product Images from "Rhein, a Natural Anthraquinone Derivative, Attenuates the Activation of Pancreatic Stellate Cells and Ameliorates Pancreatic Fibrosis in Mice with Experimental Chronic Pancreatitis"

    Article Title: Rhein, a Natural Anthraquinone Derivative, Attenuates the Activation of Pancreatic Stellate Cells and Ameliorates Pancreatic Fibrosis in Mice with Experimental Chronic Pancreatitis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0082201

    Rhein inhibits SHH and NF-κB signals in-vitro . (A) LTC-14 cells were treated with recombinant TGF-β (0, 1, 5 or 10 ng/mL) or SHH (0, 10, 50 or 100 ng/mL) in SFM for 24 hours and subjected to Western blotting analyses. Immunoblots were probed with anti-SHH antibody for the TGF-β treatment or probed with anti-α-SMA antibody for the SHH treatment. β-ACTIN was served as a loading reference. (B) LTC-14 cells were treated with TGF-β (0, 1, 5 or 10 ng/mL) in SFM for 24 hours and harvested for mRNA extraction. Transcripts of Gli1 and Gli2 were amplified by means of qPCR, normalized to the endogenous reference Gapdh and expressed as fold changes over the non-TGF-β-treated control (SFM control). * p
    Figure Legend Snippet: Rhein inhibits SHH and NF-κB signals in-vitro . (A) LTC-14 cells were treated with recombinant TGF-β (0, 1, 5 or 10 ng/mL) or SHH (0, 10, 50 or 100 ng/mL) in SFM for 24 hours and subjected to Western blotting analyses. Immunoblots were probed with anti-SHH antibody for the TGF-β treatment or probed with anti-α-SMA antibody for the SHH treatment. β-ACTIN was served as a loading reference. (B) LTC-14 cells were treated with TGF-β (0, 1, 5 or 10 ng/mL) in SFM for 24 hours and harvested for mRNA extraction. Transcripts of Gli1 and Gli2 were amplified by means of qPCR, normalized to the endogenous reference Gapdh and expressed as fold changes over the non-TGF-β-treated control (SFM control). * p

    Techniques Used: In Vitro, Recombinant, Western Blot, Amplification, Real-time Polymerase Chain Reaction

    A schematic flow chart showing the roles of TGF-β and SHH/GLI1 signaling pathways in pancreatic fibrosis.
    Figure Legend Snippet: A schematic flow chart showing the roles of TGF-β and SHH/GLI1 signaling pathways in pancreatic fibrosis.

    Techniques Used: Flow Cytometry

    Rhein suppresses SHH signaling in-vivo. (A) The fluorescent images demonstrated the immunoreactivites of GLI1 (green; FITC) and SHH (red; Rhodamine) in paraffin-embedded pancreatic tissues in which nuclei were stained blue with DAPI. (B) Protein levels of GLI1 and SHH in pancreatic homogenates of the mice were visualized on immunoblots probed with anti-GLI1 and anti-SHH antibodies whereas β-ACTIN was served as the loading reference. (C) Integrated densities of the immunobands of SHH and GLI1 were measured. Readings were normalized to the internal reference β-ACTIN and expressed as fold change over control. (D) Transcripts of Gli1 and Gli2 were amplified by means of qPCR, normalized to the endogenous reference Gapdh and expressed as fold changes over the Control group. * p
    Figure Legend Snippet: Rhein suppresses SHH signaling in-vivo. (A) The fluorescent images demonstrated the immunoreactivites of GLI1 (green; FITC) and SHH (red; Rhodamine) in paraffin-embedded pancreatic tissues in which nuclei were stained blue with DAPI. (B) Protein levels of GLI1 and SHH in pancreatic homogenates of the mice were visualized on immunoblots probed with anti-GLI1 and anti-SHH antibodies whereas β-ACTIN was served as the loading reference. (C) Integrated densities of the immunobands of SHH and GLI1 were measured. Readings were normalized to the internal reference β-ACTIN and expressed as fold change over control. (D) Transcripts of Gli1 and Gli2 were amplified by means of qPCR, normalized to the endogenous reference Gapdh and expressed as fold changes over the Control group. * p

    Techniques Used: In Vivo, Staining, Mouse Assay, Western Blot, Amplification, Real-time Polymerase Chain Reaction

    38) Product Images from "Disruption of SHH signaling cascade by SBE attenuates lung cancer progression and sensitizes DDP treatment"

    Article Title: Disruption of SHH signaling cascade by SBE attenuates lung cancer progression and sensitizes DDP treatment

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-02063-x

    Targeting SHH signaling reduces cell proliferation and clonogenicity of NSCLC via arresting cell cycle. ( A ) CCK-8 assay showing that SMO inhibitors BMS (BMS-833923) inhibited cell proliferation of both A549 and H1299 at the 2.5 µM concentration or higher. SMO inhibitors GDC (GDC-0449) significantly arrested H1299 not A549 cell growth at 5 µM concentration or above. ( B ) The representative images of colony formation of A549 and H1299 cells treated with GDC (10 µΜ), and/or BMS (5 µM), or siRNA-GLI1 from clonogenicity assay. ( C ) Flowcytometry analysis demonstrating that GDC and/or BMS significantly induced G1/S phase arrest in A549 and H1299 cells, in which additive G1/S arrest was induced with co-treatment of GDC and BMS. Data were analyzed by ModFit software and presented as mean ± SD. ( D ) Immunoblotting analysis demonstrating that GDC and/or BMS significantly reduced protein level of SMO and cell cycle regulators (Cyclin A, Cyclin B, CDK1 and CDK4) in A549 and H1299 cells, in which additive repression of SMO and those cell cycle regulators G1/S arrest was detected with co-treatment of GDC and BMS. α-Tubulin was loaded as inner control. All the co-treatments of GDC and BMS were labeled as “Combi” in this figure.
    Figure Legend Snippet: Targeting SHH signaling reduces cell proliferation and clonogenicity of NSCLC via arresting cell cycle. ( A ) CCK-8 assay showing that SMO inhibitors BMS (BMS-833923) inhibited cell proliferation of both A549 and H1299 at the 2.5 µM concentration or higher. SMO inhibitors GDC (GDC-0449) significantly arrested H1299 not A549 cell growth at 5 µM concentration or above. ( B ) The representative images of colony formation of A549 and H1299 cells treated with GDC (10 µΜ), and/or BMS (5 µM), or siRNA-GLI1 from clonogenicity assay. ( C ) Flowcytometry analysis demonstrating that GDC and/or BMS significantly induced G1/S phase arrest in A549 and H1299 cells, in which additive G1/S arrest was induced with co-treatment of GDC and BMS. Data were analyzed by ModFit software and presented as mean ± SD. ( D ) Immunoblotting analysis demonstrating that GDC and/or BMS significantly reduced protein level of SMO and cell cycle regulators (Cyclin A, Cyclin B, CDK1 and CDK4) in A549 and H1299 cells, in which additive repression of SMO and those cell cycle regulators G1/S arrest was detected with co-treatment of GDC and BMS. α-Tubulin was loaded as inner control. All the co-treatments of GDC and BMS were labeled as “Combi” in this figure.

    Techniques Used: CCK-8 Assay, Concentration Assay, Software, Labeling

    Downregulation of SMO magnifies inhibitory effects of SBE on NSCLC cell proliferation. ( A ) Proliferation assay indicating that GDC or BMS and various dose of SBE cooperatively repress NSCLC cell proliferation. ( B ) Proliferation assay showing that low dose of SBE sensitized anti-proliferative effects of SMO inhibitor (GDC and BMS) on A549 and H1299 cells. ( C ) Immunoblotting analysis demonstrating that SBE enhanced GDC or BMS-induced repression of GLI1, SMO, and PTCH on protein level.
    Figure Legend Snippet: Downregulation of SMO magnifies inhibitory effects of SBE on NSCLC cell proliferation. ( A ) Proliferation assay indicating that GDC or BMS and various dose of SBE cooperatively repress NSCLC cell proliferation. ( B ) Proliferation assay showing that low dose of SBE sensitized anti-proliferative effects of SMO inhibitor (GDC and BMS) on A549 and H1299 cells. ( C ) Immunoblotting analysis demonstrating that SBE enhanced GDC or BMS-induced repression of GLI1, SMO, and PTCH on protein level.

    Techniques Used: Proliferation Assay

    Aberrant activation of SHH signaling in lung tumors from patients with adverse prognosis. ( A ) RT-PCR analysis of the endogenous mRNA levels of SHH, SMO and GLI1 in human lung cancers relative to paired normal lung tissues. GAPDH was amplified in parallel as inner control. N and T represent normal and tumor specimens separately. ( B ) Immunoblotting analysis of the endogenous protein levels of SHH, SMO and GLI1 in human lung cancers relative to paired normal lung tissues. α-Tubulin was loaded as inner control. ( C ) Kaplan Meier overall survival (OS) curves of SMO (left, n = 1926, p = 2.2E-06 by log-rank test for significance), GLI1(middle, n = 1926, p = 0.54 by log-rank test for significance) and SHH (right, n = 1926, p = 0.23 by log-rank test for significance). ( D ) Kaplan Meier progression free survival (PFS) curves of SMO (left, n = 982, p = 1.2E-07 by log-rank test for significance), GLI1(middle, n = 982, p = 0.04 by log-rank test for significance) and SHH (right, n = 982, p = 0.022 by log-rank test for significance).
    Figure Legend Snippet: Aberrant activation of SHH signaling in lung tumors from patients with adverse prognosis. ( A ) RT-PCR analysis of the endogenous mRNA levels of SHH, SMO and GLI1 in human lung cancers relative to paired normal lung tissues. GAPDH was amplified in parallel as inner control. N and T represent normal and tumor specimens separately. ( B ) Immunoblotting analysis of the endogenous protein levels of SHH, SMO and GLI1 in human lung cancers relative to paired normal lung tissues. α-Tubulin was loaded as inner control. ( C ) Kaplan Meier overall survival (OS) curves of SMO (left, n = 1926, p = 2.2E-06 by log-rank test for significance), GLI1(middle, n = 1926, p = 0.54 by log-rank test for significance) and SHH (right, n = 1926, p = 0.23 by log-rank test for significance). ( D ) Kaplan Meier progression free survival (PFS) curves of SMO (left, n = 982, p = 1.2E-07 by log-rank test for significance), GLI1(middle, n = 982, p = 0.04 by log-rank test for significance) and SHH (right, n = 982, p = 0.022 by log-rank test for significance).

    Techniques Used: Activation Assay, Reverse Transcription Polymerase Chain Reaction, Amplification

    Up-regulation of SMO resists SBE-induced reduction in invasive growth of NSCLC cells. ( A ) Immunoblotting analysis illustrating that SMO agonist Purmorphamine (PUR) activated SMO expression in A549 and H1299 cells. ( B ) CompuSyn Software-assisted analysis of Dose-Effect curve (left) and Combination Index Plot (right) for SBE, PUR, or both showing. SBE was analyzed at dose from 26.7 to 135 µg/ml and PUR was tested at dose from 1.25 to 20 µM. ( C ) Wound healing assay showing that PUR antagonized SBE-induced repression in invasive growth of A549 and H1299 cells. ( D ) Schematic diagram depicting the SBE-mediated selective targeting inhibition of SHH-cell cycle signal axis in lung cancer therapy. As indicated in this diagram, SBE directly and selectively targets SMO both transcriptionally and translationally causing significant inhibition of GLI1 and its downstream targets including SHH signaling components and cell cycle regulators. All the co-treatments of SBE and PUR were labeled as “Combi” in this figure.
    Figure Legend Snippet: Up-regulation of SMO resists SBE-induced reduction in invasive growth of NSCLC cells. ( A ) Immunoblotting analysis illustrating that SMO agonist Purmorphamine (PUR) activated SMO expression in A549 and H1299 cells. ( B ) CompuSyn Software-assisted analysis of Dose-Effect curve (left) and Combination Index Plot (right) for SBE, PUR, or both showing. SBE was analyzed at dose from 26.7 to 135 µg/ml and PUR was tested at dose from 1.25 to 20 µM. ( C ) Wound healing assay showing that PUR antagonized SBE-induced repression in invasive growth of A549 and H1299 cells. ( D ) Schematic diagram depicting the SBE-mediated selective targeting inhibition of SHH-cell cycle signal axis in lung cancer therapy. As indicated in this diagram, SBE directly and selectively targets SMO both transcriptionally and translationally causing significant inhibition of GLI1 and its downstream targets including SHH signaling components and cell cycle regulators. All the co-treatments of SBE and PUR were labeled as “Combi” in this figure.

    Techniques Used: Expressing, Software, Wound Healing Assay, Inhibition, Labeling

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    Western Blot:

    Article Title: Pseudomonas aeruginosa-mannose-sensitive hemagglutinin inhibits chemical-induced skin cancer through suppressing hedgehog signaling
    Article Snippet: .. To determine whether the effects of TPA were mediated by Hh signaling, we detected the expression levels of SHH, SMO, and Gli-1 using Western blot. ..

    Article Title: Pseudomonas aeruginosa-mannose-sensitive hemagglutinin inhibits chemical-induced skin cancer through suppressing hedgehog signaling
    Article Snippet: .. Moreover, Western blot analysis indicated that SHH with PAM (107 and 108 cfu/mL), SMO with PAM (108 cfu/mL), or Gli-1 with PAM (107 –109 cfu/mL) in the cells was significantly decreased than those in the normal cells ( p < 0.05, ). ..

    Translocation Assay:

    Article Title: Pseudomonas aeruginosa-mannose-sensitive hemagglutinin inhibits chemical-induced skin cancer through suppressing hedgehog signaling
    Article Snippet: .. PAM (108 cfu/mL) also inhibited the protein expression of Gli-1 and its translocation, which could be abrogated by administration of 1 µM of purmorphamine ( ). ..

    In Vivo:

    Article Title: Pseudomonas aeruginosa-mannose-sensitive hemagglutinin inhibits chemical-induced skin cancer through suppressing hedgehog signaling
    Article Snippet: .. In vivo , pretreatment with PAM (108 cfu/mL) can reduce the number of tumor induced by DMBA/TPA and the epidermal thickness and inhibit the expression of Gli-1, indicating PAM inhibited EMT in vitro and in vivo , which is consistent with previous report. ..

    In Vitro:

    Article Title: Pseudomonas aeruginosa-mannose-sensitive hemagglutinin inhibits chemical-induced skin cancer through suppressing hedgehog signaling
    Article Snippet: .. In vivo , pretreatment with PAM (108 cfu/mL) can reduce the number of tumor induced by DMBA/TPA and the epidermal thickness and inhibit the expression of Gli-1, indicating PAM inhibited EMT in vitro and in vivo , which is consistent with previous report. ..

    other:

    Article Title: Pseudomonas aeruginosa-mannose-sensitive hemagglutinin inhibits chemical-induced skin cancer through suppressing hedgehog signaling
    Article Snippet: The integral optical density values of PCNA and Gli-1 were measured by CMIAS-8 color pathological image analysis system.

    Activity Assay:

    Article Title: MTOR promotes basal cell carcinoma growth through atypical PKC
    Article Snippet: .. To further delineate mTor’s mode of action, we assayed that status of aPKC, a Gli1 kinase that is necessary for high sustained Gli1 activity[ ] . mTor has been shown to phosphorylate and activate aPKC at residue T560 in mouse embryonic fibroblasts[ ] , while aPKC phosphorylates and activates Gli1 at residue T304 [ , ] . ..

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    Santa Cruz Biotechnology gli1
    Tumor cells competent for Hh signaling and OPN expression are efficient at inducing osteoblast differentiation. Stable silencing of OPN (OPNi) or <t>GLI1</t> (KD2 and KO1) from SUM1315 and MDA-MB-435 tumor cells significantly reduces the expression of ( A ) BSP [Relative to SUM1315, SUM1315-OPNi (∧p = 0.008) and KD2 (∧p = 0.0004) show lower BSP; Relative to MDA-MB-435, 435-OPNi and KO1 have decreased BSP (∧p
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    Tumor cells competent for Hh signaling and OPN expression are efficient at inducing osteoblast differentiation. Stable silencing of OPN (OPNi) or GLI1 (KD2 and KO1) from SUM1315 and MDA-MB-435 tumor cells significantly reduces the expression of ( A ) BSP [Relative to SUM1315, SUM1315-OPNi (∧p = 0.008) and KD2 (∧p = 0.0004) show lower BSP; Relative to MDA-MB-435, 435-OPNi and KO1 have decreased BSP (∧p

    Journal: PLoS ONE

    Article Title: Hedgehog Signaling in Tumor Cells Facilitates Osteoblast-Enhanced Osteolytic Metastases

    doi: 10.1371/journal.pone.0034374

    Figure Lengend Snippet: Tumor cells competent for Hh signaling and OPN expression are efficient at inducing osteoblast differentiation. Stable silencing of OPN (OPNi) or GLI1 (KD2 and KO1) from SUM1315 and MDA-MB-435 tumor cells significantly reduces the expression of ( A ) BSP [Relative to SUM1315, SUM1315-OPNi (∧p = 0.008) and KD2 (∧p = 0.0004) show lower BSP; Relative to MDA-MB-435, 435-OPNi and KO1 have decreased BSP (∧p

    Article Snippet: Treatment with the Hh inhibitor, cyclopamine, keeps GLI1 sequestered in the cytosolic compartment ( & C ) simultaneous with a reduction in the levels of OPN transcript levels (p < 0.0001) , total OPN protein expression ( ) and secreted OPN ( ) in the pre-osteoblasts.

    Techniques: Expressing, Multiple Displacement Amplification

    Active Hh signaling in the tumor cells is vital to their ability to cause osteolysis. MDA-MB-435 tumor cells were injected into the left cardiac ventricle of athymic mice; mice were euthanized 4–6 weeks later and radiographically imaged and assessed for osteolysis at the tibio-femoral junction. As represented in the Table, cells that were silenced for GLI1 expression showed an attenuated ability of osteolysis. The percent incidence of osteolysis is depicted in the adjacent graph.

    Journal: PLoS ONE

    Article Title: Hedgehog Signaling in Tumor Cells Facilitates Osteoblast-Enhanced Osteolytic Metastases

    doi: 10.1371/journal.pone.0034374

    Figure Lengend Snippet: Active Hh signaling in the tumor cells is vital to their ability to cause osteolysis. MDA-MB-435 tumor cells were injected into the left cardiac ventricle of athymic mice; mice were euthanized 4–6 weeks later and radiographically imaged and assessed for osteolysis at the tibio-femoral junction. As represented in the Table, cells that were silenced for GLI1 expression showed an attenuated ability of osteolysis. The percent incidence of osteolysis is depicted in the adjacent graph.

    Article Snippet: Treatment with the Hh inhibitor, cyclopamine, keeps GLI1 sequestered in the cytosolic compartment ( & C ) simultaneous with a reduction in the levels of OPN transcript levels (p < 0.0001) , total OPN protein expression ( ) and secreted OPN ( ) in the pre-osteoblasts.

    Techniques: Multiple Displacement Amplification, Injection, Mouse Assay, Expressing

    The Hh pathway is activated in breast tumors. Breast cancer tissues (n = 75) and normal breast tissues (n = 9) were immunohistochemically stained for ( A ) IHH and ( B ) GLI1 expression. The intensity of cytoplasmic staining was quantitated with computer-assisted image analysis in a Dako ACIS III Image Analysis System (Glostrup, Denmark). Staining intensities were recorded and represented as a scatter plot. The staining intensities indicating expression levels of IHH and GLI1 were significantly greater (p

    Journal: PLoS ONE

    Article Title: Hedgehog Signaling in Tumor Cells Facilitates Osteoblast-Enhanced Osteolytic Metastases

    doi: 10.1371/journal.pone.0034374

    Figure Lengend Snippet: The Hh pathway is activated in breast tumors. Breast cancer tissues (n = 75) and normal breast tissues (n = 9) were immunohistochemically stained for ( A ) IHH and ( B ) GLI1 expression. The intensity of cytoplasmic staining was quantitated with computer-assisted image analysis in a Dako ACIS III Image Analysis System (Glostrup, Denmark). Staining intensities were recorded and represented as a scatter plot. The staining intensities indicating expression levels of IHH and GLI1 were significantly greater (p

    Article Snippet: Treatment with the Hh inhibitor, cyclopamine, keeps GLI1 sequestered in the cytosolic compartment ( & C ) simultaneous with a reduction in the levels of OPN transcript levels (p < 0.0001) , total OPN protein expression ( ) and secreted OPN ( ) in the pre-osteoblasts.

    Techniques: Staining, Expressing

    Gli1 expression reversely correlates with E-Cadherin expression in lung SCC. (A) Expressions of Gli1 and E-Cadherin (E-Cad) in three representative tissue specimens in the UCSF cohort with Gli1 expression at a low level (upper panels) and high levels (middle and lower panels). (B) Expressions of Gli1, E-Cad and β-Catenin (β-Cat) in three representative tissue specimens in the Tianjin cohort with Gli1 expression at a low level (upper panels), a mixed expression pattern (middle panels) and a high level (lower panels). (C) Correlations between Gli1, EMT markers, and recurrence/metastasis. Statistical analysis was performed between Gli1 and E-Cad, Gli1 and β-Cat, Gli1 and recurrence/ metastasis. (D) Gli1 and E-Cad expression in four lung SCC cell lines by Western blots.

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Hedgehog/Gli promotes epithelial-mesenchymal transition in lung squamous cell carcinomas

    doi: 10.1186/1756-9966-33-34

    Figure Lengend Snippet: Gli1 expression reversely correlates with E-Cadherin expression in lung SCC. (A) Expressions of Gli1 and E-Cadherin (E-Cad) in three representative tissue specimens in the UCSF cohort with Gli1 expression at a low level (upper panels) and high levels (middle and lower panels). (B) Expressions of Gli1, E-Cad and β-Catenin (β-Cat) in three representative tissue specimens in the Tianjin cohort with Gli1 expression at a low level (upper panels), a mixed expression pattern (middle panels) and a high level (lower panels). (C) Correlations between Gli1, EMT markers, and recurrence/metastasis. Statistical analysis was performed between Gli1 and E-Cad, Gli1 and β-Cat, Gli1 and recurrence/ metastasis. (D) Gli1 and E-Cad expression in four lung SCC cell lines by Western blots.

    Article Snippet: Moreover, our analysis revealed that Gli1 significantly correlated with recurrence and metastasis of lung SCC in the Tianjin cohort (p = 0.033; Figure C).

    Techniques: Expressing, Western Blot

    Aberrant activation of the Shh pathway in lung SCC. (A) Representative protein expression of Shh, Smo, Ptch1 and Gli1 by IHC staining, scored as 0 (negative), 1 (mild positive), 2 (positive), and 3 (strong positive). (B) Expression distributions of Shh, Smo, Ptch1 and Gli1 in 177 patient tissue specimens in the Tianjin cohort. (C) Association between the expressions of Hh pathway components. Kendall’s tau-b statsitcs was used to determine the correlation between proteins. The correlation coefficient r and p values were presented in (C) . Kappa test was also performed with IHC scores of 1–3 grouped as “+”, 0 as “-”. Kappa test’s p values were labeled with*.

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Hedgehog/Gli promotes epithelial-mesenchymal transition in lung squamous cell carcinomas

    doi: 10.1186/1756-9966-33-34

    Figure Lengend Snippet: Aberrant activation of the Shh pathway in lung SCC. (A) Representative protein expression of Shh, Smo, Ptch1 and Gli1 by IHC staining, scored as 0 (negative), 1 (mild positive), 2 (positive), and 3 (strong positive). (B) Expression distributions of Shh, Smo, Ptch1 and Gli1 in 177 patient tissue specimens in the Tianjin cohort. (C) Association between the expressions of Hh pathway components. Kendall’s tau-b statsitcs was used to determine the correlation between proteins. The correlation coefficient r and p values were presented in (C) . Kappa test was also performed with IHC scores of 1–3 grouped as “+”, 0 as “-”. Kappa test’s p values were labeled with*.

    Article Snippet: Moreover, our analysis revealed that Gli1 significantly correlated with recurrence and metastasis of lung SCC in the Tianjin cohort (p = 0.033; Figure C).

    Techniques: Activation Assay, Expressing, Immunohistochemistry, Staining, Labeling

    mTor inhibition suppresses murine BCC growth and aPKC activity. A) Hematoxylin and eosin staining of dorsal back skin collected from DMSO- or Everolimus-treated Ptch1 fl/fl ; Gli1 - Cre ERT2 mice. Scale bar, 50 μm. B) Quantification of total tumor size per square area (n > 250 tumors from 5 mice). tx, treatment. C) Immunofluorescent staining of DMSO- or Everolimus-treated Ptch1 fl/fl ; Gli1 - Cre ERT2 mice for the indicated markers. Scale bar, 25 μm. D) Quantification of immunostains (n = 5 tumors from 3 mice). AU, arbitrary units. Error bars represent SEM. Significance was determined by unpaired two-tailed t test. *, p

    Journal: bioRxiv

    Article Title: MTOR promotes basal cell carcinoma growth through atypical PKC

    doi: 10.1101/2020.08.24.264598

    Figure Lengend Snippet: mTor inhibition suppresses murine BCC growth and aPKC activity. A) Hematoxylin and eosin staining of dorsal back skin collected from DMSO- or Everolimus-treated Ptch1 fl/fl ; Gli1 - Cre ERT2 mice. Scale bar, 50 μm. B) Quantification of total tumor size per square area (n > 250 tumors from 5 mice). tx, treatment. C) Immunofluorescent staining of DMSO- or Everolimus-treated Ptch1 fl/fl ; Gli1 - Cre ERT2 mice for the indicated markers. Scale bar, 25 μm. D) Quantification of immunostains (n = 5 tumors from 3 mice). AU, arbitrary units. Error bars represent SEM. Significance was determined by unpaired two-tailed t test. *, p

    Article Snippet: The following antibodies were used: mTor (rabbit, Cell Signaling Technology 2983S, 1:400), Gli1 (rabbit, Santa Cruz Biotechnology sc-20687, 1:500), Gli1 p-T304 (rabbit, 1:200) [ ], Krt14 (chicken, Fisher Scientific 50-103-0174, 1:5000), aPKC (rabbit, Santa Cruz Biotechnology sc-216, 1:500), aPKC p-T410 (rabbit, Santa Cruz Biotechnology sc-12894-R, 1:200), and aPKC p-T560 (rabbit, Abcam ab62372, 1:300).

    Techniques: Inhibition, Activity Assay, Staining, Mouse Assay, Two Tailed Test

    mTor inhibition suppresses BCC cell growth but not HH signaling. A) Gli1 mRNA levels of ASZ001 cells treated with DMSO or varying concentrations of Rapamycin, OSI-027, or Everolimus (n = 3 experiments). dR, delta reporter signal normalized to passive reference dye. B-D) MTT assay of ASZ001 cells treated with B) Everolimus, C) Rapamycin, or D) OSI-027 (n = 3 experiments). Abs, absorbance. Error bars represent SEM. Significance was determined by two-way ANOVA test. *, p

    Journal: bioRxiv

    Article Title: MTOR promotes basal cell carcinoma growth through atypical PKC

    doi: 10.1101/2020.08.24.264598

    Figure Lengend Snippet: mTor inhibition suppresses BCC cell growth but not HH signaling. A) Gli1 mRNA levels of ASZ001 cells treated with DMSO or varying concentrations of Rapamycin, OSI-027, or Everolimus (n = 3 experiments). dR, delta reporter signal normalized to passive reference dye. B-D) MTT assay of ASZ001 cells treated with B) Everolimus, C) Rapamycin, or D) OSI-027 (n = 3 experiments). Abs, absorbance. Error bars represent SEM. Significance was determined by two-way ANOVA test. *, p

    Article Snippet: The following antibodies were used: mTor (rabbit, Cell Signaling Technology 2983S, 1:400), Gli1 (rabbit, Santa Cruz Biotechnology sc-20687, 1:500), Gli1 p-T304 (rabbit, 1:200) [ ], Krt14 (chicken, Fisher Scientific 50-103-0174, 1:5000), aPKC (rabbit, Santa Cruz Biotechnology sc-216, 1:500), aPKC p-T410 (rabbit, Santa Cruz Biotechnology sc-12894-R, 1:200), and aPKC p-T560 (rabbit, Abcam ab62372, 1:300).

    Techniques: Inhibition, MTT Assay

    MTOR is overexpressed in human and mouse BCC. A) Immunofluorescent staining of indicated markers in human normal epidermis and nodular BCC. Scale bar, 50 μm. B) Quantification of MTOR immunostain intensity (n = 5 different points of measurement from 4 individual samples). AU, arbitrary units. C) Immunofluorescent staining of indicated markers in normal epidermis, normal hair follicle, or BCC derived from Ptch1 fl/fl ; Gli1 - Cre ERT2 mice. Scale bar, 25 μm. D) Quantification of mTor immunostain (n = 5 different points of measurement from 5 mice). Error bars represent SEM. Significance was determined by unpaired two-tailed t test. *, p

    Journal: bioRxiv

    Article Title: MTOR promotes basal cell carcinoma growth through atypical PKC

    doi: 10.1101/2020.08.24.264598

    Figure Lengend Snippet: MTOR is overexpressed in human and mouse BCC. A) Immunofluorescent staining of indicated markers in human normal epidermis and nodular BCC. Scale bar, 50 μm. B) Quantification of MTOR immunostain intensity (n = 5 different points of measurement from 4 individual samples). AU, arbitrary units. C) Immunofluorescent staining of indicated markers in normal epidermis, normal hair follicle, or BCC derived from Ptch1 fl/fl ; Gli1 - Cre ERT2 mice. Scale bar, 25 μm. D) Quantification of mTor immunostain (n = 5 different points of measurement from 5 mice). Error bars represent SEM. Significance was determined by unpaired two-tailed t test. *, p

    Article Snippet: The following antibodies were used: mTor (rabbit, Cell Signaling Technology 2983S, 1:400), Gli1 (rabbit, Santa Cruz Biotechnology sc-20687, 1:500), Gli1 p-T304 (rabbit, 1:200) [ ], Krt14 (chicken, Fisher Scientific 50-103-0174, 1:5000), aPKC (rabbit, Santa Cruz Biotechnology sc-216, 1:500), aPKC p-T410 (rabbit, Santa Cruz Biotechnology sc-12894-R, 1:200), and aPKC p-T560 (rabbit, Abcam ab62372, 1:300).

    Techniques: Staining, Derivative Assay, Mouse Assay, Two Tailed Test

    The Hh pathway transcriptionally regulates OPN. A , levels of GLI1 and OPN are increased in primary cutaneous cancer and metastatic melanoma. Gene microarray analysis (utilizing a Human Genome U133 Plus 2.0 array from Affymetrix, Inc.) was used to compare

    Journal: The Journal of Biological Chemistry

    Article Title: The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *

    doi: 10.1074/jbc.M109.021949

    Figure Lengend Snippet: The Hh pathway transcriptionally regulates OPN. A , levels of GLI1 and OPN are increased in primary cutaneous cancer and metastatic melanoma. Gene microarray analysis (utilizing a Human Genome U133 Plus 2.0 array from Affymetrix, Inc.) was used to compare

    Article Snippet: Thus, expression of GLI1 plays a functionally important role in the malignant behavior of tumor cells.

    Techniques: Microarray

    Silencing endogenous GLI1 expression diminishes attributes of motility, invasion, migration, and proliferation and negatively impacts tumorigenicity and metastasis. A , abrogating GLI1 expression ( KO1 and KO4 ) reduced the ability of cells to move and fill

    Journal: The Journal of Biological Chemistry

    Article Title: The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *

    doi: 10.1074/jbc.M109.021949

    Figure Lengend Snippet: Silencing endogenous GLI1 expression diminishes attributes of motility, invasion, migration, and proliferation and negatively impacts tumorigenicity and metastasis. A , abrogating GLI1 expression ( KO1 and KO4 ) reduced the ability of cells to move and fill

    Article Snippet: Thus, expression of GLI1 plays a functionally important role in the malignant behavior of tumor cells.

    Techniques: Expressing, Migration

    The OPN promoter bears a GLI1-binding site that responds to GLI1. A , mutating the putative GLI1-binding site in the OPN promoter makes the OPN promoter insensitive to the effects of GLI1. The OPN promoter (+112 to −352 (pGL3-OPN-352)) was significantly

    Journal: The Journal of Biological Chemistry

    Article Title: The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *

    doi: 10.1074/jbc.M109.021949

    Figure Lengend Snippet: The OPN promoter bears a GLI1-binding site that responds to GLI1. A , mutating the putative GLI1-binding site in the OPN promoter makes the OPN promoter insensitive to the effects of GLI1. The OPN promoter (+112 to −352 (pGL3-OPN-352)) was significantly

    Article Snippet: Thus, expression of GLI1 plays a functionally important role in the malignant behavior of tumor cells.

    Techniques: Binding Assay

    Restoring the availability of OPN-initiated signaling in GLI1-silenced cells reinstates their motility and ability to migrate and invade. A , immunoblot representing the restored OPN in cells that have been stably knocked down for GLI1 (GLI1 KO; and consequently

    Journal: The Journal of Biological Chemistry

    Article Title: The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *

    doi: 10.1074/jbc.M109.021949

    Figure Lengend Snippet: Restoring the availability of OPN-initiated signaling in GLI1-silenced cells reinstates their motility and ability to migrate and invade. A , immunoblot representing the restored OPN in cells that have been stably knocked down for GLI1 (GLI1 KO; and consequently

    Article Snippet: Thus, expression of GLI1 plays a functionally important role in the malignant behavior of tumor cells.

    Techniques: Stable Transfection

    shRNA to GLI1 abrogates expression of OPN and brings about a partial reversal of EMT. A , MDA-MB-435 cells were stably transfected with shRNA to GLI1. Clones KO1 to KO4 were stably silenced for GLI1 and show notably reduced OPN expression. B , real time

    Journal: The Journal of Biological Chemistry

    Article Title: The Hedgehog Pathway Transcription Factor GLI1 Promotes Malignant Behavior of Cancer Cells by Up-regulating Osteopontin *

    doi: 10.1074/jbc.M109.021949

    Figure Lengend Snippet: shRNA to GLI1 abrogates expression of OPN and brings about a partial reversal of EMT. A , MDA-MB-435 cells were stably transfected with shRNA to GLI1. Clones KO1 to KO4 were stably silenced for GLI1 and show notably reduced OPN expression. B , real time

    Article Snippet: Thus, expression of GLI1 plays a functionally important role in the malignant behavior of tumor cells.

    Techniques: shRNA, Expressing, Multiple Displacement Amplification, Stable Transfection, Transfection, Clone Assay