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sonic hedgehog  (R&D Systems)


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    R&D Systems sonic hedgehog
    Sonic Hedgehog, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 103 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+human+shh/Recombinant+Human+Sonic+Hedgehog%2FShh+(C24II)+N-Terminus/pm41942110-57-11-14
    Average 94 stars, based on 103 article reviews
    sonic hedgehog - by Bioz Stars, 2026-09
    94/100 stars

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    Related Articles

    Recombinant:

    Article Title: miR-301a-mediated crosstalk between the Hedgehog and HIPPO/YAP signaling pathways promotes pancreatic cancer
    Article Snippet: These cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, USA) and penicillin-streptomycin solution (100 U/mL penicillin/100 ng/mL streptomycin, Hyclone, USA) at 37°C with 5% CO2. .. Recombinant human SHH, TNF-α and IL-1β were acquired from R&D Systems, USA. .. The cells were seeded in T25 culture dishes (Corning, USA).

    Article Title: Bi-allelic Variations of SMO in Humans Cause a Broad Spectrum of Developmental Anomalies Due to Abnormal Hedgehog Signaling.
    Article Snippet: .. For SHH pathway induction, either recombinant human SHH (rhSHH, R&D Systems, Cat: 8908-SH-005) or smoothened agonist (SAG, Cat: sc-202814) was used. rhSHHwas used at a final concentration of 100 mg/mL, for 24 h, after 48 h serum starvation, and SAG was used at final concentration of 10 mg/mL, for 24 h, after 48 h serum starvation. ..

    Article Title: Dysfunction of Shh signaling activates autophagy to inhibit trophoblast motility in recurrent miscarriage
    Article Snippet: .. JAR cells were seeded into 96-well plates at 1000 cells/well and cultured for 24 h. Then, the culture medium was replaced with a fresh complete growth medium plus a final concentration of 5 μM cyclopamine (Tocris, 1623, USA) or 0.5 μg/ml recombinant human Shh (R&D, 1845-SH). ..

    Article Title: Bi-allelic Variations of SMO in Humans Cause a Broad Spectrum of Developmental Anomalies Due to Abnormal Hedgehog Signaling
    Article Snippet: .. For SHH pathway induction, either recombinant human SHH (rhSHH, R&D Systems, Cat: 8908-SH-005) or smoothened agonist (SAG, Cat: sc-202814) was used. rhSHH was used at a final concentration of 100 μg/mL, for 24 h, after 48 h serum starvation, and SAG was used at final concentration of 10 μg/mL, for 24 h, after 48 h serum starvation. ..

    Article Title: LGALS3 (galectin 3) mediates an unconventional secretion of SNCA/α-synuclein in response to lysosomal membrane damage by the autophagic-lysosomal pathway in human midbrain dopamine neurons
    Article Snippet: Prior to floor plate induction, iPSCs were grown to 80–90% confluence. iPSC cultures were monitored daily with an in-hood EVOS Core XL cell imaging system (Thermo Fisher Scientific). .. iPSCs were incubated in knockout serum replacement media (KSRM; Thermo Fisher Scientific, 10828028) containing the Small Mothers Against Decapetaplegic inhibitors, LDN193189 (LDN, 100 nM; ReproCell, 04–0074) and SB431542 (SB, 10 μM; R & D System, 1614/1) for 24 h, KSRM containing LDN, SB, recombinant human SHH (sonic hedgehog signaling molecule; 100 pg/μl; R&D Systems, 1845-SH-025), purmorphamine (2 μM; ReproCell, 04–0009) and recombinant human FGF8 (fibroblast growth factor 8; 100 pg/μl; R&D Systems, 4745-F8-050) for 48 h, and KSRM containing LDN, SB, SHH, purmorphamine (Reprocell, 04–0009), FGF8, and CHIR99021 (CHIR, 3 μM; ReproCell, 04–0004) for 48 h. KSRM media was comprised of 82.20% KnockOut Dulbecco’s Modified Eagle’s Medium (Thermo Fisher Scientific, 10–829-018), 14.68% Knockout Serum Replacement (Thermo Fisher Scientific, 10–828-028), 0.98% 200 mM L-glutamine (Thermo Fisher Scientific, 35050061), 0.98% Minimum Essential Media Non-Essential Amino Acids solution (Thermo Fisher Scientific, 11140050), 0.18% 55 mM 2-mercopatoethanol, and 0.98% 10,000 U/ml penicillin-streptomycin solution (Thermo Fisher Scientific, 15–140-122). .. Next, iPSCs were incubated with a 3:1 ratio of KSRM to SM1 media containing LDN, SB, SHH, purmorphamine, FGF8, and CHIR for 48 h, a 1:1 ratio of KSRM to SM1 media containing LDN and CHIR for 48 h, and 1:3 ratio of KSRM to SM1 media containing LDN and CHIR for 48 h. SM1 media was comprised of 96.16% Neurobasal Medium (Thermo Fisher Scientific, 21103049), 1.92% NeuroCult SM1 Neuronal Supplement (Stem Cell Technologies, 05711), 0.96% 200 mM L-glutamine solution (Thermo Fisher Scientific, 35,050,061), and 0.96% 10,000 U/ml penicillin-streptomycin solution (Thermo Fisher Scientific, 15140122).

    Article Title: Dysfunction of Shh signaling activates autophagy to inhibit trophoblast motility in recurrent miscarriage
    Article Snippet: .. Cyclopamine (Tocris, 1623, USA), recombinant human Shh (R&D, 1845-SH), chloroquine (Sigma, C6628), GANT61 (Selleck Chemicals, S8075), Lipofectamine 3000 (Thermo Fisher Scientific, L3000015), Gli2 shRNA1 (5′-GUACCAUUACGAGCCUCAUUC-3′), Gli2 shRNA2 (5′-CAACGCCCCCCACCCGUAC-3′), Gli3 shRNA1 (5′-UUGAAGGUUGCACAAAGGC-3′), Gli3 shRNA2 (5′-AAGAGAUUAAACUGACUUU-3′), BECN1 shRNA1 (5′-GGATGACA GTGAACAGTTA-3′), and BECN1 shRNA2 (5′-CCCGTGGAATGGAATGAGA-3′). .. Total RNA was isolated by using TRIzol reagent (Takara, China) according to the manufacturer’s instructions.

    Article Title: miR-301a-mediated crosstalk between the Hedgehog and HIPPO/YAP signaling pathways promotes pancreatic cancer.
    Article Snippet: These cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, USA) and penicillin-streptomycin solution (100 U/mL penicillin/100 ng/mL streptomycin, Hyclone, USA) at 37°C with 5% CO2. .. Recombinant human SHH, TNF-α and IL-1β were acquired from R&D Systems, USA. .. The cells were seeded in T25 culture dishes (Corning, USA).

    Article Title: Increasing Ciliary ARL13B Expression Drives Active and Inhibitor-Resistant SMO and GLI into Glioma Primary Cilia
    Article Snippet: .. Where indicated, indicated cell lines were treated with various drugs including cyclopamine (Calbiochem; Cat#239803) (diluted in dimethyl sulfoxide (DMSO)), temozolomide (TMZ) (Sigma; Cat# T2577) (diluted in DMSO), GDC-0449 (Cayman Chemical; Cat #13613), SAG (Calbiochem; Cat #566661), recombinant human SHH (high activity (R&D Systems: Cat # 8908-SH-005/CF), and fixed after indicated treatment durations and processed as described below. .. Where indicated, cells were also transiently transfected with pFlag-Smo (PD22) (gift from G. Pazour), pCAG-Ar13b WT :GFPpb, pDest-Arl13b T35N :GFP cDNA using Lipofectamine 3000(Invitrogen; cat # L3000-008), and fixed at the indicated timepoints as described below.

    Concentration Assay:

    Article Title: Bi-allelic Variations of SMO in Humans Cause a Broad Spectrum of Developmental Anomalies Due to Abnormal Hedgehog Signaling.
    Article Snippet: .. For SHH pathway induction, either recombinant human SHH (rhSHH, R&D Systems, Cat: 8908-SH-005) or smoothened agonist (SAG, Cat: sc-202814) was used. rhSHHwas used at a final concentration of 100 mg/mL, for 24 h, after 48 h serum starvation, and SAG was used at final concentration of 10 mg/mL, for 24 h, after 48 h serum starvation. ..

    Article Title: Dysfunction of Shh signaling activates autophagy to inhibit trophoblast motility in recurrent miscarriage
    Article Snippet: .. JAR cells were seeded into 96-well plates at 1000 cells/well and cultured for 24 h. Then, the culture medium was replaced with a fresh complete growth medium plus a final concentration of 5 μM cyclopamine (Tocris, 1623, USA) or 0.5 μg/ml recombinant human Shh (R&D, 1845-SH). ..

    Article Title: Bi-allelic Variations of SMO in Humans Cause a Broad Spectrum of Developmental Anomalies Due to Abnormal Hedgehog Signaling
    Article Snippet: .. For SHH pathway induction, either recombinant human SHH (rhSHH, R&D Systems, Cat: 8908-SH-005) or smoothened agonist (SAG, Cat: sc-202814) was used. rhSHH was used at a final concentration of 100 μg/mL, for 24 h, after 48 h serum starvation, and SAG was used at final concentration of 10 μg/mL, for 24 h, after 48 h serum starvation. ..

    Cell Culture:

    Article Title: Dysfunction of Shh signaling activates autophagy to inhibit trophoblast motility in recurrent miscarriage
    Article Snippet: .. JAR cells were seeded into 96-well plates at 1000 cells/well and cultured for 24 h. Then, the culture medium was replaced with a fresh complete growth medium plus a final concentration of 5 μM cyclopamine (Tocris, 1623, USA) or 0.5 μg/ml recombinant human Shh (R&D, 1845-SH). ..

    Incubation:

    Article Title: LGALS3 (galectin 3) mediates an unconventional secretion of SNCA/α-synuclein in response to lysosomal membrane damage by the autophagic-lysosomal pathway in human midbrain dopamine neurons
    Article Snippet: Prior to floor plate induction, iPSCs were grown to 80–90% confluence. iPSC cultures were monitored daily with an in-hood EVOS Core XL cell imaging system (Thermo Fisher Scientific). .. iPSCs were incubated in knockout serum replacement media (KSRM; Thermo Fisher Scientific, 10828028) containing the Small Mothers Against Decapetaplegic inhibitors, LDN193189 (LDN, 100 nM; ReproCell, 04–0074) and SB431542 (SB, 10 μM; R & D System, 1614/1) for 24 h, KSRM containing LDN, SB, recombinant human SHH (sonic hedgehog signaling molecule; 100 pg/μl; R&D Systems, 1845-SH-025), purmorphamine (2 μM; ReproCell, 04–0009) and recombinant human FGF8 (fibroblast growth factor 8; 100 pg/μl; R&D Systems, 4745-F8-050) for 48 h, and KSRM containing LDN, SB, SHH, purmorphamine (Reprocell, 04–0009), FGF8, and CHIR99021 (CHIR, 3 μM; ReproCell, 04–0004) for 48 h. KSRM media was comprised of 82.20% KnockOut Dulbecco’s Modified Eagle’s Medium (Thermo Fisher Scientific, 10–829-018), 14.68% Knockout Serum Replacement (Thermo Fisher Scientific, 10–828-028), 0.98% 200 mM L-glutamine (Thermo Fisher Scientific, 35050061), 0.98% Minimum Essential Media Non-Essential Amino Acids solution (Thermo Fisher Scientific, 11140050), 0.18% 55 mM 2-mercopatoethanol, and 0.98% 10,000 U/ml penicillin-streptomycin solution (Thermo Fisher Scientific, 15–140-122). .. Next, iPSCs were incubated with a 3:1 ratio of KSRM to SM1 media containing LDN, SB, SHH, purmorphamine, FGF8, and CHIR for 48 h, a 1:1 ratio of KSRM to SM1 media containing LDN and CHIR for 48 h, and 1:3 ratio of KSRM to SM1 media containing LDN and CHIR for 48 h. SM1 media was comprised of 96.16% Neurobasal Medium (Thermo Fisher Scientific, 21103049), 1.92% NeuroCult SM1 Neuronal Supplement (Stem Cell Technologies, 05711), 0.96% 200 mM L-glutamine solution (Thermo Fisher Scientific, 35,050,061), and 0.96% 10,000 U/ml penicillin-streptomycin solution (Thermo Fisher Scientific, 15140122).

    Knock-Out:

    Article Title: LGALS3 (galectin 3) mediates an unconventional secretion of SNCA/α-synuclein in response to lysosomal membrane damage by the autophagic-lysosomal pathway in human midbrain dopamine neurons
    Article Snippet: Prior to floor plate induction, iPSCs were grown to 80–90% confluence. iPSC cultures were monitored daily with an in-hood EVOS Core XL cell imaging system (Thermo Fisher Scientific). .. iPSCs were incubated in knockout serum replacement media (KSRM; Thermo Fisher Scientific, 10828028) containing the Small Mothers Against Decapetaplegic inhibitors, LDN193189 (LDN, 100 nM; ReproCell, 04–0074) and SB431542 (SB, 10 μM; R & D System, 1614/1) for 24 h, KSRM containing LDN, SB, recombinant human SHH (sonic hedgehog signaling molecule; 100 pg/μl; R&D Systems, 1845-SH-025), purmorphamine (2 μM; ReproCell, 04–0009) and recombinant human FGF8 (fibroblast growth factor 8; 100 pg/μl; R&D Systems, 4745-F8-050) for 48 h, and KSRM containing LDN, SB, SHH, purmorphamine (Reprocell, 04–0009), FGF8, and CHIR99021 (CHIR, 3 μM; ReproCell, 04–0004) for 48 h. KSRM media was comprised of 82.20% KnockOut Dulbecco’s Modified Eagle’s Medium (Thermo Fisher Scientific, 10–829-018), 14.68% Knockout Serum Replacement (Thermo Fisher Scientific, 10–828-028), 0.98% 200 mM L-glutamine (Thermo Fisher Scientific, 35050061), 0.98% Minimum Essential Media Non-Essential Amino Acids solution (Thermo Fisher Scientific, 11140050), 0.18% 55 mM 2-mercopatoethanol, and 0.98% 10,000 U/ml penicillin-streptomycin solution (Thermo Fisher Scientific, 15–140-122). .. Next, iPSCs were incubated with a 3:1 ratio of KSRM to SM1 media containing LDN, SB, SHH, purmorphamine, FGF8, and CHIR for 48 h, a 1:1 ratio of KSRM to SM1 media containing LDN and CHIR for 48 h, and 1:3 ratio of KSRM to SM1 media containing LDN and CHIR for 48 h. SM1 media was comprised of 96.16% Neurobasal Medium (Thermo Fisher Scientific, 21103049), 1.92% NeuroCult SM1 Neuronal Supplement (Stem Cell Technologies, 05711), 0.96% 200 mM L-glutamine solution (Thermo Fisher Scientific, 35,050,061), and 0.96% 10,000 U/ml penicillin-streptomycin solution (Thermo Fisher Scientific, 15140122).

    Modification:

    Article Title: LGALS3 (galectin 3) mediates an unconventional secretion of SNCA/α-synuclein in response to lysosomal membrane damage by the autophagic-lysosomal pathway in human midbrain dopamine neurons
    Article Snippet: Prior to floor plate induction, iPSCs were grown to 80–90% confluence. iPSC cultures were monitored daily with an in-hood EVOS Core XL cell imaging system (Thermo Fisher Scientific). .. iPSCs were incubated in knockout serum replacement media (KSRM; Thermo Fisher Scientific, 10828028) containing the Small Mothers Against Decapetaplegic inhibitors, LDN193189 (LDN, 100 nM; ReproCell, 04–0074) and SB431542 (SB, 10 μM; R & D System, 1614/1) for 24 h, KSRM containing LDN, SB, recombinant human SHH (sonic hedgehog signaling molecule; 100 pg/μl; R&D Systems, 1845-SH-025), purmorphamine (2 μM; ReproCell, 04–0009) and recombinant human FGF8 (fibroblast growth factor 8; 100 pg/μl; R&D Systems, 4745-F8-050) for 48 h, and KSRM containing LDN, SB, SHH, purmorphamine (Reprocell, 04–0009), FGF8, and CHIR99021 (CHIR, 3 μM; ReproCell, 04–0004) for 48 h. KSRM media was comprised of 82.20% KnockOut Dulbecco’s Modified Eagle’s Medium (Thermo Fisher Scientific, 10–829-018), 14.68% Knockout Serum Replacement (Thermo Fisher Scientific, 10–828-028), 0.98% 200 mM L-glutamine (Thermo Fisher Scientific, 35050061), 0.98% Minimum Essential Media Non-Essential Amino Acids solution (Thermo Fisher Scientific, 11140050), 0.18% 55 mM 2-mercopatoethanol, and 0.98% 10,000 U/ml penicillin-streptomycin solution (Thermo Fisher Scientific, 15–140-122). .. Next, iPSCs were incubated with a 3:1 ratio of KSRM to SM1 media containing LDN, SB, SHH, purmorphamine, FGF8, and CHIR for 48 h, a 1:1 ratio of KSRM to SM1 media containing LDN and CHIR for 48 h, and 1:3 ratio of KSRM to SM1 media containing LDN and CHIR for 48 h. SM1 media was comprised of 96.16% Neurobasal Medium (Thermo Fisher Scientific, 21103049), 1.92% NeuroCult SM1 Neuronal Supplement (Stem Cell Technologies, 05711), 0.96% 200 mM L-glutamine solution (Thermo Fisher Scientific, 35,050,061), and 0.96% 10,000 U/ml penicillin-streptomycin solution (Thermo Fisher Scientific, 15140122).

    Activity Assay:

    Article Title: Increasing Ciliary ARL13B Expression Drives Active and Inhibitor-Resistant SMO and GLI into Glioma Primary Cilia
    Article Snippet: .. Where indicated, indicated cell lines were treated with various drugs including cyclopamine (Calbiochem; Cat#239803) (diluted in dimethyl sulfoxide (DMSO)), temozolomide (TMZ) (Sigma; Cat# T2577) (diluted in DMSO), GDC-0449 (Cayman Chemical; Cat #13613), SAG (Calbiochem; Cat #566661), recombinant human SHH (high activity (R&D Systems: Cat # 8908-SH-005/CF), and fixed after indicated treatment durations and processed as described below. .. Where indicated, cells were also transiently transfected with pFlag-Smo (PD22) (gift from G. Pazour), pCAG-Ar13b WT :GFPpb, pDest-Arl13b T35N :GFP cDNA using Lipofectamine 3000(Invitrogen; cat # L3000-008), and fixed at the indicated timepoints as described below.



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    Image Search Results


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

    Journal: Molecular Cancer

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

    doi: 10.1186/s12943-026-02611-y

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

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

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

    Loss of Men1 in enteric glial cells stimulates GLI1/2-dependent transcriptional reprogramming. A Combined fluorescence and phase contrast images of 5-day-old primary enteric glial cell (EGC) cultures from Sox10-CreER T2 ; LSL-tdTomato mice and CreER T2 negative controls. Top panel shows TdTomato+ EGCs after 48 h exposure to 4-hydroxytamoxifen 4-OHT (2 µM). B TdTomato + EGCs were sorted by FACS to enrich for a pure SOX10 + cell population. C Combined fluorescence and phase contrast images of FACS-enriched SOX10-tdTomato + EGCs. D Fluctuations in HH pathway mRNA levels were evaluated in SOX10-tdTomato + EGCs 72 h following siRNA-mediated Men1 silencing. siRNA treatment consisted of four pooled siRNAs targeting the Men1 gene ( si -Men1 , 25 nM) or non-targeting (si-NT, 25 nM) controls. ( n = 5). E Immunofluorescence images of SHH expression in si-NT and si-Men1 treated EGCs (SHH = red pseudo-color, DAPI = blue). Inset shows higher power image. F Western blot analysis of si-NT and si-Men1 EGCs after 72 h treatment. SHH-FL = 55 kDa full length peptide; SHH- N = 22 kDa N-terminal peptide. ( n = 3). G Quantitation of protein expression in panel (F) normalized to GAPDH loading control. ( n = 3). H Relative fold-change in glial lineage transcripts and ( I ) neuroendocrine and neural progenitor transcripts in si-NT and si-Men1 treated EGCs. ( n = 6). J qPCR analysis of HH pathway genes and ( K ) neuroendocrine and neural progenitor transcriptsin si-NT and si-Men1 EGCs after 72 h treatment with GANT61 (10 µM) or vismodegib (VISMO 20 µM). ( n = 3). For all plots, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by Two-way ANOVA with Sidak post-test. L Immunofluorescence images of si-NT and si-Men1 EGCs after 96 h siRNA knockdown and 72 h treatment with vehicle or GANT61. Menin = green, GFAP = magenta, SHH = yellow. M Significant GSEA pathways in enteric glial cells following 5-days of si- Men1 knockdown compared to non-targeting control. GSEA was performed on Men1 -depleted cells and cells co-treated with ( N ) si- Men1 , si- Gli1 , and si- Gli2 , and ( O ) cells co-treated with si- Men1 and GANT61 (10 µM). P – R KEGG pathway enrichment analysis comparing the same groups as shown in panels ( M – O ). S Heatmap showing significant DEGs mapped to the cell cycle, HH signaling, epigenetic regulation, neural stem cell (NSC) reprogramming, neuronal and neuroendocrine differentiation

    Journal: Molecular Cancer

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

    doi: 10.1186/s12943-026-02611-y

    Figure Lengend Snippet: Loss of Men1 in enteric glial cells stimulates GLI1/2-dependent transcriptional reprogramming. A Combined fluorescence and phase contrast images of 5-day-old primary enteric glial cell (EGC) cultures from Sox10-CreER T2 ; LSL-tdTomato mice and CreER T2 negative controls. Top panel shows TdTomato+ EGCs after 48 h exposure to 4-hydroxytamoxifen 4-OHT (2 µM). B TdTomato + EGCs were sorted by FACS to enrich for a pure SOX10 + cell population. C Combined fluorescence and phase contrast images of FACS-enriched SOX10-tdTomato + EGCs. D Fluctuations in HH pathway mRNA levels were evaluated in SOX10-tdTomato + EGCs 72 h following siRNA-mediated Men1 silencing. siRNA treatment consisted of four pooled siRNAs targeting the Men1 gene ( si -Men1 , 25 nM) or non-targeting (si-NT, 25 nM) controls. ( n = 5). E Immunofluorescence images of SHH expression in si-NT and si-Men1 treated EGCs (SHH = red pseudo-color, DAPI = blue). Inset shows higher power image. F Western blot analysis of si-NT and si-Men1 EGCs after 72 h treatment. SHH-FL = 55 kDa full length peptide; SHH- N = 22 kDa N-terminal peptide. ( n = 3). G Quantitation of protein expression in panel (F) normalized to GAPDH loading control. ( n = 3). H Relative fold-change in glial lineage transcripts and ( I ) neuroendocrine and neural progenitor transcripts in si-NT and si-Men1 treated EGCs. ( n = 6). J qPCR analysis of HH pathway genes and ( K ) neuroendocrine and neural progenitor transcriptsin si-NT and si-Men1 EGCs after 72 h treatment with GANT61 (10 µM) or vismodegib (VISMO 20 µM). ( n = 3). For all plots, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by Two-way ANOVA with Sidak post-test. L Immunofluorescence images of si-NT and si-Men1 EGCs after 96 h siRNA knockdown and 72 h treatment with vehicle or GANT61. Menin = green, GFAP = magenta, SHH = yellow. M Significant GSEA pathways in enteric glial cells following 5-days of si- Men1 knockdown compared to non-targeting control. GSEA was performed on Men1 -depleted cells and cells co-treated with ( N ) si- Men1 , si- Gli1 , and si- Gli2 , and ( O ) cells co-treated with si- Men1 and GANT61 (10 µM). P – R KEGG pathway enrichment analysis comparing the same groups as shown in panels ( M – O ). S Heatmap showing significant DEGs mapped to the cell cycle, HH signaling, epigenetic regulation, neural stem cell (NSC) reprogramming, neuronal and neuroendocrine differentiation

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

    Techniques: Fluorescence, Immunofluorescence, Expressing, Western Blot, Quantitation Assay, Control, Knockdown