sag Search Results


sag  (Tocris)
95
Tocris sag
Sag, supplied by Tocris, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/sag/bio_rxiv__2024__11__26__624368-276-73-74?v=Tocris
Average 95 stars, based on 1 article reviews
sag - by Bioz Stars, 2026-08
95/100 stars
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sag 21k  (Tocris)
94
Tocris sag 21k
A) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with Vismodegib + PD173074 for 72 hours, in the presence or absence of WNT agonist (CHIR99021, 1-6 μM) or WNT inhibitor (XAV939, 1 μM). This revealed that 2 μM of CHIR99021 (indicated by white circle) was optimal for inducing dorsal forebrain marker EMX2 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . HH inh.: HEDGEHOG inhibitor. FGFR inh.: FGF receptor inhibitor. B) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of FGF2 (20 ng/mL) or FGFR inhibitor (PD173074, 100 nM). This revealed that FGF inhibition enhanced dorsal forebrain markers EMX2 and PAX6 , while preventing erroneous WNT-induced expression of midbrain marker EN1 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . FGFRi: FGF receptor inhibitor. C) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of BMP4 (10 ng/mL) or BMPR inhibitors (DMH1 [250 nM] or LDN193189 [100 nM]). Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. D) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist <t>21K</t> (0.1-5 nM) for 72 hours to generate ventral forebrain. This showed that high HEDGEHOG pathway activation, together with WNT inhibition, led to maximum expression of ventral forebrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. HH: Hedgehog pathway agonist. WNTi: WNT inhibitor. E) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG inhibitor (Vismodegib, 150 nM) and WNT agonist (CHIR99021, 2 μM) for 72 hours to generate dorsal forebrain, in the presence or absence of BMP agonist (BMP4, 10 ng/mL) or BMPR inhibitor (LDN193189, 100 nM). This showed that BMP inhibition promoted expression of dorsal forebrain markers GSX2 and PAX3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. F) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG pathway inhibitor Vismodegib (150 nM) in the presence or absence of increasing doses of WNT pathway agonist CHIR99021 (1-4 μM) for 72 hours to generate dorsal forebrain. This showed that moderate WNT pathway activation, together with HH inhibition, led to maximum expression of dorsal hindbrain markers GSX2 and OLIG3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. G) scRNAseq of H7 hPSC-derived day-4 hindbrain progenitors, and day-7 dorsal hindbrain or ventral hindbrain progenitors. Colors denote differentiation conditions. An asterisk (*) indicates that the legend is identical to that shown in , and is reproduced here to aid the interpretation of other images shown in this subpanel. H) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with either Vismodegib (150 nM) + CHIR99021 (2 μM) + DMH1 (250 nM) for 72 hours to generate dorsal forebrain, or alternatively, 21K (5 nM) + XAV939 (1 μM) for 72 hours to generate ventral forebrain. Gene expression is shown relative to the sample with the highest expression in this experiment. I) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with WNT pathway inhibitor XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (50 pM-5 nM) for 72 hours to generate ventral hindbrain. This showed that high doses of HEDGEHOG agonist, together with WNT inhibitor, were required to maximally induce ventral hindbrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. WNT inh.: WNT inhibitor. qPCR data depicts the mean of two biological replicates, with s.e.m. shown. In vitro scRNAseq data is from a single experiment.
Sag 21k, supplied by Tocris, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/sag/bio_rxiv__2025__07__02__662771-283-17-21?v=Tocris
Average 94 stars, based on 1 article reviews
sag 21k - by Bioz Stars, 2026-08
94/100 stars
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sag1 s7779  (Selleck Chemicals)
94
Selleck Chemicals sag1 s7779
A) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with Vismodegib + PD173074 for 72 hours, in the presence or absence of WNT agonist (CHIR99021, 1-6 μM) or WNT inhibitor (XAV939, 1 μM). This revealed that 2 μM of CHIR99021 (indicated by white circle) was optimal for inducing dorsal forebrain marker EMX2 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . HH inh.: HEDGEHOG inhibitor. FGFR inh.: FGF receptor inhibitor. B) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of FGF2 (20 ng/mL) or FGFR inhibitor (PD173074, 100 nM). This revealed that FGF inhibition enhanced dorsal forebrain markers EMX2 and PAX6 , while preventing erroneous WNT-induced expression of midbrain marker EN1 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . FGFRi: FGF receptor inhibitor. C) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of BMP4 (10 ng/mL) or BMPR inhibitors (DMH1 [250 nM] or LDN193189 [100 nM]). Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. D) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist <t>21K</t> (0.1-5 nM) for 72 hours to generate ventral forebrain. This showed that high HEDGEHOG pathway activation, together with WNT inhibition, led to maximum expression of ventral forebrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. HH: Hedgehog pathway agonist. WNTi: WNT inhibitor. E) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG inhibitor (Vismodegib, 150 nM) and WNT agonist (CHIR99021, 2 μM) for 72 hours to generate dorsal forebrain, in the presence or absence of BMP agonist (BMP4, 10 ng/mL) or BMPR inhibitor (LDN193189, 100 nM). This showed that BMP inhibition promoted expression of dorsal forebrain markers GSX2 and PAX3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. F) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG pathway inhibitor Vismodegib (150 nM) in the presence or absence of increasing doses of WNT pathway agonist CHIR99021 (1-4 μM) for 72 hours to generate dorsal forebrain. This showed that moderate WNT pathway activation, together with HH inhibition, led to maximum expression of dorsal hindbrain markers GSX2 and OLIG3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. G) scRNAseq of H7 hPSC-derived day-4 hindbrain progenitors, and day-7 dorsal hindbrain or ventral hindbrain progenitors. Colors denote differentiation conditions. An asterisk (*) indicates that the legend is identical to that shown in , and is reproduced here to aid the interpretation of other images shown in this subpanel. H) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with either Vismodegib (150 nM) + CHIR99021 (2 μM) + DMH1 (250 nM) for 72 hours to generate dorsal forebrain, or alternatively, 21K (5 nM) + XAV939 (1 μM) for 72 hours to generate ventral forebrain. Gene expression is shown relative to the sample with the highest expression in this experiment. I) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with WNT pathway inhibitor XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (50 pM-5 nM) for 72 hours to generate ventral hindbrain. This showed that high doses of HEDGEHOG agonist, together with WNT inhibitor, were required to maximally induce ventral hindbrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. WNT inh.: WNT inhibitor. qPCR data depicts the mean of two biological replicates, with s.e.m. shown. In vitro scRNAseq data is from a single experiment.
Sag1 S7779, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/sag/pmc10937112__mmc3-703-6-11?v=Selleck+Chemicals
Average 94 stars, based on 1 article reviews
sag1 s7779 - by Bioz Stars, 2026-08
94/100 stars
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transfer vector  (OriGene)
91
OriGene transfer vector
A) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with Vismodegib + PD173074 for 72 hours, in the presence or absence of WNT agonist (CHIR99021, 1-6 μM) or WNT inhibitor (XAV939, 1 μM). This revealed that 2 μM of CHIR99021 (indicated by white circle) was optimal for inducing dorsal forebrain marker EMX2 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . HH inh.: HEDGEHOG inhibitor. FGFR inh.: FGF receptor inhibitor. B) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of FGF2 (20 ng/mL) or FGFR inhibitor (PD173074, 100 nM). This revealed that FGF inhibition enhanced dorsal forebrain markers EMX2 and PAX6 , while preventing erroneous WNT-induced expression of midbrain marker EN1 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . FGFRi: FGF receptor inhibitor. C) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of BMP4 (10 ng/mL) or BMPR inhibitors (DMH1 [250 nM] or LDN193189 [100 nM]). Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. D) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist <t>21K</t> (0.1-5 nM) for 72 hours to generate ventral forebrain. This showed that high HEDGEHOG pathway activation, together with WNT inhibition, led to maximum expression of ventral forebrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. HH: Hedgehog pathway agonist. WNTi: WNT inhibitor. E) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG inhibitor (Vismodegib, 150 nM) and WNT agonist (CHIR99021, 2 μM) for 72 hours to generate dorsal forebrain, in the presence or absence of BMP agonist (BMP4, 10 ng/mL) or BMPR inhibitor (LDN193189, 100 nM). This showed that BMP inhibition promoted expression of dorsal forebrain markers GSX2 and PAX3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. F) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG pathway inhibitor Vismodegib (150 nM) in the presence or absence of increasing doses of WNT pathway agonist CHIR99021 (1-4 μM) for 72 hours to generate dorsal forebrain. This showed that moderate WNT pathway activation, together with HH inhibition, led to maximum expression of dorsal hindbrain markers GSX2 and OLIG3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. G) scRNAseq of H7 hPSC-derived day-4 hindbrain progenitors, and day-7 dorsal hindbrain or ventral hindbrain progenitors. Colors denote differentiation conditions. An asterisk (*) indicates that the legend is identical to that shown in , and is reproduced here to aid the interpretation of other images shown in this subpanel. H) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with either Vismodegib (150 nM) + CHIR99021 (2 μM) + DMH1 (250 nM) for 72 hours to generate dorsal forebrain, or alternatively, 21K (5 nM) + XAV939 (1 μM) for 72 hours to generate ventral forebrain. Gene expression is shown relative to the sample with the highest expression in this experiment. I) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with WNT pathway inhibitor XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (50 pM-5 nM) for 72 hours to generate ventral hindbrain. This showed that high doses of HEDGEHOG agonist, together with WNT inhibitor, were required to maximally induce ventral hindbrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. WNT inh.: WNT inhibitor. qPCR data depicts the mean of two biological replicates, with s.e.m. shown. In vitro scRNAseq data is from a single experiment.
Transfer Vector, supplied by OriGene, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/sag/pm36581221-56-38-49?v=OriGene
Average 91 stars, based on 1 article reviews
transfer vector - by Bioz Stars, 2026-08
91/100 stars
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53299 rrid ab 629595  (Santa Cruz Biotechnology)
93
Santa Cruz Biotechnology 53299 rrid ab 629595
A) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with Vismodegib + PD173074 for 72 hours, in the presence or absence of WNT agonist (CHIR99021, 1-6 μM) or WNT inhibitor (XAV939, 1 μM). This revealed that 2 μM of CHIR99021 (indicated by white circle) was optimal for inducing dorsal forebrain marker EMX2 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . HH inh.: HEDGEHOG inhibitor. FGFR inh.: FGF receptor inhibitor. B) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of FGF2 (20 ng/mL) or FGFR inhibitor (PD173074, 100 nM). This revealed that FGF inhibition enhanced dorsal forebrain markers EMX2 and PAX6 , while preventing erroneous WNT-induced expression of midbrain marker EN1 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . FGFRi: FGF receptor inhibitor. C) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of BMP4 (10 ng/mL) or BMPR inhibitors (DMH1 [250 nM] or LDN193189 [100 nM]). Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. D) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist <t>21K</t> (0.1-5 nM) for 72 hours to generate ventral forebrain. This showed that high HEDGEHOG pathway activation, together with WNT inhibition, led to maximum expression of ventral forebrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. HH: Hedgehog pathway agonist. WNTi: WNT inhibitor. E) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG inhibitor (Vismodegib, 150 nM) and WNT agonist (CHIR99021, 2 μM) for 72 hours to generate dorsal forebrain, in the presence or absence of BMP agonist (BMP4, 10 ng/mL) or BMPR inhibitor (LDN193189, 100 nM). This showed that BMP inhibition promoted expression of dorsal forebrain markers GSX2 and PAX3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. F) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG pathway inhibitor Vismodegib (150 nM) in the presence or absence of increasing doses of WNT pathway agonist CHIR99021 (1-4 μM) for 72 hours to generate dorsal forebrain. This showed that moderate WNT pathway activation, together with HH inhibition, led to maximum expression of dorsal hindbrain markers GSX2 and OLIG3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. G) scRNAseq of H7 hPSC-derived day-4 hindbrain progenitors, and day-7 dorsal hindbrain or ventral hindbrain progenitors. Colors denote differentiation conditions. An asterisk (*) indicates that the legend is identical to that shown in , and is reproduced here to aid the interpretation of other images shown in this subpanel. H) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with either Vismodegib (150 nM) + CHIR99021 (2 μM) + DMH1 (250 nM) for 72 hours to generate dorsal forebrain, or alternatively, 21K (5 nM) + XAV939 (1 μM) for 72 hours to generate ventral forebrain. Gene expression is shown relative to the sample with the highest expression in this experiment. I) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with WNT pathway inhibitor XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (50 pM-5 nM) for 72 hours to generate ventral hindbrain. This showed that high doses of HEDGEHOG agonist, together with WNT inhibitor, were required to maximally induce ventral hindbrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. WNT inh.: WNT inhibitor. qPCR data depicts the mean of two biological replicates, with s.e.m. shown. In vitro scRNAseq data is from a single experiment.
53299 Rrid Ab 629595, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/sag/pmc12890720-11-4-2?v=Santa+Cruz+Biotechnology
Average 93 stars, based on 1 article reviews
53299 rrid ab 629595 - by Bioz Stars, 2026-08
93/100 stars
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smo agonist sag dihydrochloride  (Tocris)
91
Tocris smo agonist sag dihydrochloride
A) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with Vismodegib + PD173074 for 72 hours, in the presence or absence of WNT agonist (CHIR99021, 1-6 μM) or WNT inhibitor (XAV939, 1 μM). This revealed that 2 μM of CHIR99021 (indicated by white circle) was optimal for inducing dorsal forebrain marker EMX2 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . HH inh.: HEDGEHOG inhibitor. FGFR inh.: FGF receptor inhibitor. B) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of FGF2 (20 ng/mL) or FGFR inhibitor (PD173074, 100 nM). This revealed that FGF inhibition enhanced dorsal forebrain markers EMX2 and PAX6 , while preventing erroneous WNT-induced expression of midbrain marker EN1 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . FGFRi: FGF receptor inhibitor. C) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of BMP4 (10 ng/mL) or BMPR inhibitors (DMH1 [250 nM] or LDN193189 [100 nM]). Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. D) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist <t>21K</t> (0.1-5 nM) for 72 hours to generate ventral forebrain. This showed that high HEDGEHOG pathway activation, together with WNT inhibition, led to maximum expression of ventral forebrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. HH: Hedgehog pathway agonist. WNTi: WNT inhibitor. E) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG inhibitor (Vismodegib, 150 nM) and WNT agonist (CHIR99021, 2 μM) for 72 hours to generate dorsal forebrain, in the presence or absence of BMP agonist (BMP4, 10 ng/mL) or BMPR inhibitor (LDN193189, 100 nM). This showed that BMP inhibition promoted expression of dorsal forebrain markers GSX2 and PAX3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. F) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG pathway inhibitor Vismodegib (150 nM) in the presence or absence of increasing doses of WNT pathway agonist CHIR99021 (1-4 μM) for 72 hours to generate dorsal forebrain. This showed that moderate WNT pathway activation, together with HH inhibition, led to maximum expression of dorsal hindbrain markers GSX2 and OLIG3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. G) scRNAseq of H7 hPSC-derived day-4 hindbrain progenitors, and day-7 dorsal hindbrain or ventral hindbrain progenitors. Colors denote differentiation conditions. An asterisk (*) indicates that the legend is identical to that shown in , and is reproduced here to aid the interpretation of other images shown in this subpanel. H) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with either Vismodegib (150 nM) + CHIR99021 (2 μM) + DMH1 (250 nM) for 72 hours to generate dorsal forebrain, or alternatively, 21K (5 nM) + XAV939 (1 μM) for 72 hours to generate ventral forebrain. Gene expression is shown relative to the sample with the highest expression in this experiment. I) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with WNT pathway inhibitor XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (50 pM-5 nM) for 72 hours to generate ventral hindbrain. This showed that high doses of HEDGEHOG agonist, together with WNT inhibitor, were required to maximally induce ventral hindbrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. WNT inh.: WNT inhibitor. qPCR data depicts the mean of two biological replicates, with s.e.m. shown. In vitro scRNAseq data is from a single experiment.
Smo Agonist Sag Dihydrochloride, supplied by Tocris, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/sag/pmc07112150-118-37-42?v=Tocris
Average 91 stars, based on 1 article reviews
smo agonist sag dihydrochloride - by Bioz Stars, 2026-08
91/100 stars
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agonist sag  (Santa Cruz Biotechnology)
93
Santa Cruz Biotechnology agonist sag
A) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with Vismodegib + PD173074 for 72 hours, in the presence or absence of WNT agonist (CHIR99021, 1-6 μM) or WNT inhibitor (XAV939, 1 μM). This revealed that 2 μM of CHIR99021 (indicated by white circle) was optimal for inducing dorsal forebrain marker EMX2 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . HH inh.: HEDGEHOG inhibitor. FGFR inh.: FGF receptor inhibitor. B) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of FGF2 (20 ng/mL) or FGFR inhibitor (PD173074, 100 nM). This revealed that FGF inhibition enhanced dorsal forebrain markers EMX2 and PAX6 , while preventing erroneous WNT-induced expression of midbrain marker EN1 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . FGFRi: FGF receptor inhibitor. C) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of BMP4 (10 ng/mL) or BMPR inhibitors (DMH1 [250 nM] or LDN193189 [100 nM]). Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. D) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist <t>21K</t> (0.1-5 nM) for 72 hours to generate ventral forebrain. This showed that high HEDGEHOG pathway activation, together with WNT inhibition, led to maximum expression of ventral forebrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. HH: Hedgehog pathway agonist. WNTi: WNT inhibitor. E) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG inhibitor (Vismodegib, 150 nM) and WNT agonist (CHIR99021, 2 μM) for 72 hours to generate dorsal forebrain, in the presence or absence of BMP agonist (BMP4, 10 ng/mL) or BMPR inhibitor (LDN193189, 100 nM). This showed that BMP inhibition promoted expression of dorsal forebrain markers GSX2 and PAX3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. F) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG pathway inhibitor Vismodegib (150 nM) in the presence or absence of increasing doses of WNT pathway agonist CHIR99021 (1-4 μM) for 72 hours to generate dorsal forebrain. This showed that moderate WNT pathway activation, together with HH inhibition, led to maximum expression of dorsal hindbrain markers GSX2 and OLIG3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. G) scRNAseq of H7 hPSC-derived day-4 hindbrain progenitors, and day-7 dorsal hindbrain or ventral hindbrain progenitors. Colors denote differentiation conditions. An asterisk (*) indicates that the legend is identical to that shown in , and is reproduced here to aid the interpretation of other images shown in this subpanel. H) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with either Vismodegib (150 nM) + CHIR99021 (2 μM) + DMH1 (250 nM) for 72 hours to generate dorsal forebrain, or alternatively, 21K (5 nM) + XAV939 (1 μM) for 72 hours to generate ventral forebrain. Gene expression is shown relative to the sample with the highest expression in this experiment. I) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with WNT pathway inhibitor XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (50 pM-5 nM) for 72 hours to generate ventral hindbrain. This showed that high doses of HEDGEHOG agonist, together with WNT inhibitor, were required to maximally induce ventral hindbrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. WNT inh.: WNT inhibitor. qPCR data depicts the mean of two biological replicates, with s.e.m. shown. In vitro scRNAseq data is from a single experiment.
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75 287 arrestin sag  (OriGene)
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A) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with Vismodegib + PD173074 for 72 hours, in the presence or absence of WNT agonist (CHIR99021, 1-6 μM) or WNT inhibitor (XAV939, 1 μM). This revealed that 2 μM of CHIR99021 (indicated by white circle) was optimal for inducing dorsal forebrain marker EMX2 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . HH inh.: HEDGEHOG inhibitor. FGFR inh.: FGF receptor inhibitor. B) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of FGF2 (20 ng/mL) or FGFR inhibitor (PD173074, 100 nM). This revealed that FGF inhibition enhanced dorsal forebrain markers EMX2 and PAX6 , while preventing erroneous WNT-induced expression of midbrain marker EN1 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . FGFRi: FGF receptor inhibitor. C) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of BMP4 (10 ng/mL) or BMPR inhibitors (DMH1 [250 nM] or LDN193189 [100 nM]). Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. D) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist <t>21K</t> (0.1-5 nM) for 72 hours to generate ventral forebrain. This showed that high HEDGEHOG pathway activation, together with WNT inhibition, led to maximum expression of ventral forebrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. HH: Hedgehog pathway agonist. WNTi: WNT inhibitor. E) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG inhibitor (Vismodegib, 150 nM) and WNT agonist (CHIR99021, 2 μM) for 72 hours to generate dorsal forebrain, in the presence or absence of BMP agonist (BMP4, 10 ng/mL) or BMPR inhibitor (LDN193189, 100 nM). This showed that BMP inhibition promoted expression of dorsal forebrain markers GSX2 and PAX3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. F) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG pathway inhibitor Vismodegib (150 nM) in the presence or absence of increasing doses of WNT pathway agonist CHIR99021 (1-4 μM) for 72 hours to generate dorsal forebrain. This showed that moderate WNT pathway activation, together with HH inhibition, led to maximum expression of dorsal hindbrain markers GSX2 and OLIG3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. G) scRNAseq of H7 hPSC-derived day-4 hindbrain progenitors, and day-7 dorsal hindbrain or ventral hindbrain progenitors. Colors denote differentiation conditions. An asterisk (*) indicates that the legend is identical to that shown in , and is reproduced here to aid the interpretation of other images shown in this subpanel. H) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with either Vismodegib (150 nM) + CHIR99021 (2 μM) + DMH1 (250 nM) for 72 hours to generate dorsal forebrain, or alternatively, 21K (5 nM) + XAV939 (1 μM) for 72 hours to generate ventral forebrain. Gene expression is shown relative to the sample with the highest expression in this experiment. I) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with WNT pathway inhibitor XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (50 pM-5 nM) for 72 hours to generate ventral hindbrain. This showed that high doses of HEDGEHOG agonist, together with WNT inhibitor, were required to maximally induce ventral hindbrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. WNT inh.: WNT inhibitor. qPCR data depicts the mean of two biological replicates, with s.e.m. shown. In vitro scRNAseq data is from a single experiment.
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DMBT1 is down-regulated in HNSCC, and overexpression of DMBT1 suppresses invasion and tumor progression. (A) Meta-analysis showing down-regulation of DMBT1 in HNSCC relative to normal tissue. The datasets and accession numbers are given in . (B) Down-regulation of DMBT1 is correlated with poor overall survival in patients with HNSCC. The analyses were performed with the Rickman Head-Neck dataset (reporter ID, 208250_s_at; Oncomine; n = 81; *, P < 0.05; log-rank test). (C) DMBT1 gene expression in HNSCC cell lines. Total RNA from immortalized (HOK16B), primary keratinocytes (HOK5973), and HNSCC cell lines UM-SCC-(1, 14A, 29, 47, 81B, 104) was used to generate cDNAs, and Q-RT-PCR was performed. Data were analyzed by the relative quantification method with normalization to GAPDH and then relative to keratinocytes (*, P < 0.001; one-way ANOVA; error bars represent SD). Each sample was analyzed in triplicate, and the average fold-change was determined. (D) DMBT1 was stably overexpressed in UM-SCC-29 as verified by immunoblot analysis ( n = 2). (E) Overexpression of DMBT1 suppresses invasion in vitro at 48 h. Each color represents an independent experiment with three replicates in each experiment (*, P < 0.05; t test). (F) Nuclear pleomorphism is significantly less in tumors with UM-SCC-29-DMBT1 cells than in control tumors with <t>UM-SCC-29-pCMV6</t> cells (***, P < 0.001; t test; n = 9; error bars represent SD).
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DMBT1 is down-regulated in HNSCC, and overexpression of DMBT1 suppresses invasion and tumor progression. (A) Meta-analysis showing down-regulation of DMBT1 in HNSCC relative to normal tissue. The datasets and accession numbers are given in . (B) Down-regulation of DMBT1 is correlated with poor overall survival in patients with HNSCC. The analyses were performed with the Rickman Head-Neck dataset (reporter ID, 208250_s_at; Oncomine; n = 81; *, P < 0.05; log-rank test). (C) DMBT1 gene expression in HNSCC cell lines. Total RNA from immortalized (HOK16B), primary keratinocytes (HOK5973), and HNSCC cell lines UM-SCC-(1, 14A, 29, 47, 81B, 104) was used to generate cDNAs, and Q-RT-PCR was performed. Data were analyzed by the relative quantification method with normalization to GAPDH and then relative to keratinocytes (*, P < 0.001; one-way ANOVA; error bars represent SD). Each sample was analyzed in triplicate, and the average fold-change was determined. (D) DMBT1 was stably overexpressed in UM-SCC-29 as verified by immunoblot analysis ( n = 2). (E) Overexpression of DMBT1 suppresses invasion in vitro at 48 h. Each color represents an independent experiment with three replicates in each experiment (*, P < 0.05; t test). (F) Nuclear pleomorphism is significantly less in tumors with UM-SCC-29-DMBT1 cells than in control tumors with <t>UM-SCC-29-pCMV6</t> cells (***, P < 0.001; t test; n = 9; error bars represent SD).
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Image Search Results


A) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with Vismodegib + PD173074 for 72 hours, in the presence or absence of WNT agonist (CHIR99021, 1-6 μM) or WNT inhibitor (XAV939, 1 μM). This revealed that 2 μM of CHIR99021 (indicated by white circle) was optimal for inducing dorsal forebrain marker EMX2 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . HH inh.: HEDGEHOG inhibitor. FGFR inh.: FGF receptor inhibitor. B) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of FGF2 (20 ng/mL) or FGFR inhibitor (PD173074, 100 nM). This revealed that FGF inhibition enhanced dorsal forebrain markers EMX2 and PAX6 , while preventing erroneous WNT-induced expression of midbrain marker EN1 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . FGFRi: FGF receptor inhibitor. C) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of BMP4 (10 ng/mL) or BMPR inhibitors (DMH1 [250 nM] or LDN193189 [100 nM]). Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. D) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (0.1-5 nM) for 72 hours to generate ventral forebrain. This showed that high HEDGEHOG pathway activation, together with WNT inhibition, led to maximum expression of ventral forebrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. HH: Hedgehog pathway agonist. WNTi: WNT inhibitor. E) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG inhibitor (Vismodegib, 150 nM) and WNT agonist (CHIR99021, 2 μM) for 72 hours to generate dorsal forebrain, in the presence or absence of BMP agonist (BMP4, 10 ng/mL) or BMPR inhibitor (LDN193189, 100 nM). This showed that BMP inhibition promoted expression of dorsal forebrain markers GSX2 and PAX3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. F) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG pathway inhibitor Vismodegib (150 nM) in the presence or absence of increasing doses of WNT pathway agonist CHIR99021 (1-4 μM) for 72 hours to generate dorsal forebrain. This showed that moderate WNT pathway activation, together with HH inhibition, led to maximum expression of dorsal hindbrain markers GSX2 and OLIG3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. G) scRNAseq of H7 hPSC-derived day-4 hindbrain progenitors, and day-7 dorsal hindbrain or ventral hindbrain progenitors. Colors denote differentiation conditions. An asterisk (*) indicates that the legend is identical to that shown in , and is reproduced here to aid the interpretation of other images shown in this subpanel. H) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with either Vismodegib (150 nM) + CHIR99021 (2 μM) + DMH1 (250 nM) for 72 hours to generate dorsal forebrain, or alternatively, 21K (5 nM) + XAV939 (1 μM) for 72 hours to generate ventral forebrain. Gene expression is shown relative to the sample with the highest expression in this experiment. I) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with WNT pathway inhibitor XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (50 pM-5 nM) for 72 hours to generate ventral hindbrain. This showed that high doses of HEDGEHOG agonist, together with WNT inhibitor, were required to maximally induce ventral hindbrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. WNT inh.: WNT inhibitor. qPCR data depicts the mean of two biological replicates, with s.e.m. shown. In vitro scRNAseq data is from a single experiment.

Journal: bioRxiv

Article Title: Two parallel lineage-committed progenitors contribute to the developing brain

doi: 10.1101/2025.07.02.662771

Figure Lengend Snippet: A) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with Vismodegib + PD173074 for 72 hours, in the presence or absence of WNT agonist (CHIR99021, 1-6 μM) or WNT inhibitor (XAV939, 1 μM). This revealed that 2 μM of CHIR99021 (indicated by white circle) was optimal for inducing dorsal forebrain marker EMX2 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . HH inh.: HEDGEHOG inhibitor. FGFR inh.: FGF receptor inhibitor. B) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of FGF2 (20 ng/mL) or FGFR inhibitor (PD173074, 100 nM). This revealed that FGF inhibition enhanced dorsal forebrain markers EMX2 and PAX6 , while preventing erroneous WNT-induced expression of midbrain marker EN1 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . FGFRi: FGF receptor inhibitor. C) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with CHIR99021 (2 μM) + PD173074 (100 nM) for 72 hours, in the presence or absence of BMP4 (10 ng/mL) or BMPR inhibitors (DMH1 [250 nM] or LDN193189 [100 nM]). Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. D) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then anterior neural ectoderm within 24 hours, followed by forebrain progenitors within 48 hours. Forebrain progenitors were then treated with XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (0.1-5 nM) for 72 hours to generate ventral forebrain. This showed that high HEDGEHOG pathway activation, together with WNT inhibition, led to maximum expression of ventral forebrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. HH: Hedgehog pathway agonist. WNTi: WNT inhibitor. E) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG inhibitor (Vismodegib, 150 nM) and WNT agonist (CHIR99021, 2 μM) for 72 hours to generate dorsal forebrain, in the presence or absence of BMP agonist (BMP4, 10 ng/mL) or BMPR inhibitor (LDN193189, 100 nM). This showed that BMP inhibition promoted expression of dorsal forebrain markers GSX2 and PAX3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. F) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with HEDGEHOG pathway inhibitor Vismodegib (150 nM) in the presence or absence of increasing doses of WNT pathway agonist CHIR99021 (1-4 μM) for 72 hours to generate dorsal forebrain. This showed that moderate WNT pathway activation, together with HH inhibition, led to maximum expression of dorsal hindbrain markers GSX2 and OLIG3 . Gene expression is shown relative to reference gene YWHAZ ; 100% indicates that a gene is expressed at the same level as YWHAZ . BMPRi: BMP receptor inhibitor. G) scRNAseq of H7 hPSC-derived day-4 hindbrain progenitors, and day-7 dorsal hindbrain or ventral hindbrain progenitors. Colors denote differentiation conditions. An asterisk (*) indicates that the legend is identical to that shown in , and is reproduced here to aid the interpretation of other images shown in this subpanel. H) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with either Vismodegib (150 nM) + CHIR99021 (2 μM) + DMH1 (250 nM) for 72 hours to generate dorsal forebrain, or alternatively, 21K (5 nM) + XAV939 (1 μM) for 72 hours to generate ventral forebrain. Gene expression is shown relative to the sample with the highest expression in this experiment. I) H1 hPSCs were differentiated into definitive ectoderm within 24 hours, and then posterior neural ectoderm within 24 hours, followed by hindbrain progenitors within 48 hours. Hindbrain progenitors were then treated with WNT pathway inhibitor XAV939 (1 μM) in the presence or absence of increasing doses of HEDGEHOG pathway agonist 21K (50 pM-5 nM) for 72 hours to generate ventral hindbrain. This showed that high doses of HEDGEHOG agonist, together with WNT inhibitor, were required to maximally induce ventral hindbrain markers. Gene expression is shown relative to the sample with the highest expression in this experiment. WNT inh.: WNT inhibitor. qPCR data depicts the mean of two biological replicates, with s.e.m. shown. In vitro scRNAseq data is from a single experiment.

Article Snippet: D5, D6, and D7 dorsal forebrain media consisted of CDM2 supplemented with XAV939 (1 μM, Tocris) and SAG 21K (5 nM, Tocris).

Techniques: Marker, Gene Expression, Inhibition, Expressing, Activation Assay, Derivative Assay, In Vitro

DMBT1 is down-regulated in HNSCC, and overexpression of DMBT1 suppresses invasion and tumor progression. (A) Meta-analysis showing down-regulation of DMBT1 in HNSCC relative to normal tissue. The datasets and accession numbers are given in . (B) Down-regulation of DMBT1 is correlated with poor overall survival in patients with HNSCC. The analyses were performed with the Rickman Head-Neck dataset (reporter ID, 208250_s_at; Oncomine; n = 81; *, P < 0.05; log-rank test). (C) DMBT1 gene expression in HNSCC cell lines. Total RNA from immortalized (HOK16B), primary keratinocytes (HOK5973), and HNSCC cell lines UM-SCC-(1, 14A, 29, 47, 81B, 104) was used to generate cDNAs, and Q-RT-PCR was performed. Data were analyzed by the relative quantification method with normalization to GAPDH and then relative to keratinocytes (*, P < 0.001; one-way ANOVA; error bars represent SD). Each sample was analyzed in triplicate, and the average fold-change was determined. (D) DMBT1 was stably overexpressed in UM-SCC-29 as verified by immunoblot analysis ( n = 2). (E) Overexpression of DMBT1 suppresses invasion in vitro at 48 h. Each color represents an independent experiment with three replicates in each experiment (*, P < 0.05; t test). (F) Nuclear pleomorphism is significantly less in tumors with UM-SCC-29-DMBT1 cells than in control tumors with UM-SCC-29-pCMV6 cells (***, P < 0.001; t test; n = 9; error bars represent SD).

Journal: The Journal of Experimental Medicine

Article Title: Squamous cell carcinoma subverts adjacent histologically normal epithelium to promote lateral invasion

doi: 10.1084/jem.20200944

Figure Lengend Snippet: DMBT1 is down-regulated in HNSCC, and overexpression of DMBT1 suppresses invasion and tumor progression. (A) Meta-analysis showing down-regulation of DMBT1 in HNSCC relative to normal tissue. The datasets and accession numbers are given in . (B) Down-regulation of DMBT1 is correlated with poor overall survival in patients with HNSCC. The analyses were performed with the Rickman Head-Neck dataset (reporter ID, 208250_s_at; Oncomine; n = 81; *, P < 0.05; log-rank test). (C) DMBT1 gene expression in HNSCC cell lines. Total RNA from immortalized (HOK16B), primary keratinocytes (HOK5973), and HNSCC cell lines UM-SCC-(1, 14A, 29, 47, 81B, 104) was used to generate cDNAs, and Q-RT-PCR was performed. Data were analyzed by the relative quantification method with normalization to GAPDH and then relative to keratinocytes (*, P < 0.001; one-way ANOVA; error bars represent SD). Each sample was analyzed in triplicate, and the average fold-change was determined. (D) DMBT1 was stably overexpressed in UM-SCC-29 as verified by immunoblot analysis ( n = 2). (E) Overexpression of DMBT1 suppresses invasion in vitro at 48 h. Each color represents an independent experiment with three replicates in each experiment (*, P < 0.05; t test). (F) Nuclear pleomorphism is significantly less in tumors with UM-SCC-29-DMBT1 cells than in control tumors with UM-SCC-29-pCMV6 cells (***, P < 0.001; t test; n = 9; error bars represent SD).

Article Snippet: UM-SCC-1 and UM-SCC-29 cells were transfected with empty vector (pCMV6) or DMBT1 plasmids (#RC215454; OriGene).

Techniques: Over Expression, Gene Expression, Reverse Transcription Polymerase Chain Reaction, Quantitative Proteomics, Stable Transfection, Western Blot, In Vitro, Control

DMBT1 inhibits invasion and metastasis. (A) Down-regulation of DMBT1 is associated with distant metastasis-free survival. The analysis was performed with the Rickman Head-Neck dataset (reporter ID, 208250_s_at; Oncomine; n = 81; *, P < 0.05; log-rank test). (B) Protein lysates from immortalized (HOK16B), primary keratinocytes (HOK5973), and HNSCC cell lines UM-SCC-(1, 14A, 29, 47, 81B, 104) were immunoblotted with anti-DMBT1 antibody. Signal intensity was quantified by densitometric analysis with normalization to actin (loading control) and then expressed as percent of the signal intensity in HOK16B ( n = 2). (C) Overexpression of DMBT1 inhibits tumor growth in the CAM in vivo model. UM-SCC-29-pCMV6 and UM-SCC-29-DMBT1 cells (1 × 10 6 ) were seeded on the chick CAM (*, P < 0.05; n = 6; t test). (D) Overexpression of DMBT1 suppresses invasion in the CAM assay (left scale bar = 1 mm; right scale bar = 500 µm). Tumor sections from the CAM were stained with H&E. Arrows point to invasive islands. (E) Invasive tumor islands in tumor sections were plotted as the total number of invasive islands per sample (*, P < 0.05; t test). (F) Metastases (Mets) to the lower CAM were quantified with quantitative Alu-PCR (*, P < 0.05; t test). (G) UM-SCC-29 overexpressing DMBT1 (UM-SCC-29-DMBT1) or empty vector (UM-SCC-29-pCMV6; 1 × 10 6 ) were injected subcutaneously in athymic nude mice. Days 0 and 7 denote the time of injection and the start of tumor measurements, respectively. (H) Tumor size for mice injected with UM-SCC-29-pCMV6 or UM-SCC-29-DMBT1 cells (****, P < 0.0001; n = 9/group; linear mixed model of tumor volume with random effect for mouse and fixed effects for time). (I) Quantification of mitoses from 10 high power fields (HPF) in each mouse tissue section (images at 40× magnification; *, P < 0.05; t test). (J) H&E-stained images for pCMV6 and DMBT1 groups. Representative images show the predominant pattern of invasion. Arrows denote tumor islands at the invasive front (outside scale bar = 200 µm; inside scale bar = 40 µm). (K) Predominant pattern of invasion analyzed in tissue sections from each mouse (**, P < 0.01; t test).

Journal: The Journal of Experimental Medicine

Article Title: Squamous cell carcinoma subverts adjacent histologically normal epithelium to promote lateral invasion

doi: 10.1084/jem.20200944

Figure Lengend Snippet: DMBT1 inhibits invasion and metastasis. (A) Down-regulation of DMBT1 is associated with distant metastasis-free survival. The analysis was performed with the Rickman Head-Neck dataset (reporter ID, 208250_s_at; Oncomine; n = 81; *, P < 0.05; log-rank test). (B) Protein lysates from immortalized (HOK16B), primary keratinocytes (HOK5973), and HNSCC cell lines UM-SCC-(1, 14A, 29, 47, 81B, 104) were immunoblotted with anti-DMBT1 antibody. Signal intensity was quantified by densitometric analysis with normalization to actin (loading control) and then expressed as percent of the signal intensity in HOK16B ( n = 2). (C) Overexpression of DMBT1 inhibits tumor growth in the CAM in vivo model. UM-SCC-29-pCMV6 and UM-SCC-29-DMBT1 cells (1 × 10 6 ) were seeded on the chick CAM (*, P < 0.05; n = 6; t test). (D) Overexpression of DMBT1 suppresses invasion in the CAM assay (left scale bar = 1 mm; right scale bar = 500 µm). Tumor sections from the CAM were stained with H&E. Arrows point to invasive islands. (E) Invasive tumor islands in tumor sections were plotted as the total number of invasive islands per sample (*, P < 0.05; t test). (F) Metastases (Mets) to the lower CAM were quantified with quantitative Alu-PCR (*, P < 0.05; t test). (G) UM-SCC-29 overexpressing DMBT1 (UM-SCC-29-DMBT1) or empty vector (UM-SCC-29-pCMV6; 1 × 10 6 ) were injected subcutaneously in athymic nude mice. Days 0 and 7 denote the time of injection and the start of tumor measurements, respectively. (H) Tumor size for mice injected with UM-SCC-29-pCMV6 or UM-SCC-29-DMBT1 cells (****, P < 0.0001; n = 9/group; linear mixed model of tumor volume with random effect for mouse and fixed effects for time). (I) Quantification of mitoses from 10 high power fields (HPF) in each mouse tissue section (images at 40× magnification; *, P < 0.05; t test). (J) H&E-stained images for pCMV6 and DMBT1 groups. Representative images show the predominant pattern of invasion. Arrows denote tumor islands at the invasive front (outside scale bar = 200 µm; inside scale bar = 40 µm). (K) Predominant pattern of invasion analyzed in tissue sections from each mouse (**, P < 0.01; t test).

Article Snippet: UM-SCC-1 and UM-SCC-29 cells were transfected with empty vector (pCMV6) or DMBT1 plasmids (#RC215454; OriGene).

Techniques: Control, Over Expression, In Vivo, Chick Chorioallantoic Membrane Assay, Staining, Plasmid Preparation, Injection

DMBT1 regulates mRNA stability and transcription of MMP9 and E-cadherin via ZEB1. (A) Immunoblot for EMT markers in UM-SCC-29-pCMV6 and UM-SCC-29-DMBT1 cells ( n = 2). (B–E) DMBT1 inhibits invasion via up-regulation of E-cadherin (E-cad). UM-SCC-29-DMBT1 cells were transiently transfected with siNT or siE-cad-1 (B and C) or siE-cad-2 (D and E). (B and D) Immunoblots verify suppression of E-cadherin. (C and E) Invasion assays. Scatter plots show three independent experiments; each color represents one experiment with at least duplicates in each (**, P < 0.001; ***, P < 0.0001; one-way ANOVA; error bars represent SD). (F and G) Overexpression of MMP9 and ZEB1 rescue invasion. UM-SCC-29-DMBT1 cells were transduced with empty vector (pLX304) or MMP9 or ZEB1. UM-SCC-29-pCMV was transduced with pLX304 as a control for UM-SCC-29-DMBT1-pLX304. Lysates were immunoblotted with anti-MMP9, anti-ZEB1, and anti-GAPDH (F). Invasion was quantified at 48 h after seeding (G). Scatter plots show two independent experiments; each color represents an independent experiment with three replicates in each (****, P < 0.0001; one-way ANOVA; error bars represent SD). (H and I) UM-SCC-29-pCMV6 and UM-SCC-29-DMBT1 were incubated with actinomycin D (1 µg/ml), and RNA was isolated at the indicated time. MMP-9 (H) and E-cadherin (I) transcripts, quantified by Q-RT-PCR, were normalized to GAPDH. Results are presented as percent of corresponding transcript at time 0. Values are mean ± SEM ( n = 3). (J and K) Normalized luciferase activity of MMP9 (J) and CDH1 (K) promoters in UM-SCC-29-pCMV6 and UM-SCC-29-DMBT1. Scatter plots show three independent experiments; each color represents an independent experiment (***, P < 0.001; t test; error bars represent SD).

Journal: The Journal of Experimental Medicine

Article Title: Squamous cell carcinoma subverts adjacent histologically normal epithelium to promote lateral invasion

doi: 10.1084/jem.20200944

Figure Lengend Snippet: DMBT1 regulates mRNA stability and transcription of MMP9 and E-cadherin via ZEB1. (A) Immunoblot for EMT markers in UM-SCC-29-pCMV6 and UM-SCC-29-DMBT1 cells ( n = 2). (B–E) DMBT1 inhibits invasion via up-regulation of E-cadherin (E-cad). UM-SCC-29-DMBT1 cells were transiently transfected with siNT or siE-cad-1 (B and C) or siE-cad-2 (D and E). (B and D) Immunoblots verify suppression of E-cadherin. (C and E) Invasion assays. Scatter plots show three independent experiments; each color represents one experiment with at least duplicates in each (**, P < 0.001; ***, P < 0.0001; one-way ANOVA; error bars represent SD). (F and G) Overexpression of MMP9 and ZEB1 rescue invasion. UM-SCC-29-DMBT1 cells were transduced with empty vector (pLX304) or MMP9 or ZEB1. UM-SCC-29-pCMV was transduced with pLX304 as a control for UM-SCC-29-DMBT1-pLX304. Lysates were immunoblotted with anti-MMP9, anti-ZEB1, and anti-GAPDH (F). Invasion was quantified at 48 h after seeding (G). Scatter plots show two independent experiments; each color represents an independent experiment with three replicates in each (****, P < 0.0001; one-way ANOVA; error bars represent SD). (H and I) UM-SCC-29-pCMV6 and UM-SCC-29-DMBT1 were incubated with actinomycin D (1 µg/ml), and RNA was isolated at the indicated time. MMP-9 (H) and E-cadherin (I) transcripts, quantified by Q-RT-PCR, were normalized to GAPDH. Results are presented as percent of corresponding transcript at time 0. Values are mean ± SEM ( n = 3). (J and K) Normalized luciferase activity of MMP9 (J) and CDH1 (K) promoters in UM-SCC-29-pCMV6 and UM-SCC-29-DMBT1. Scatter plots show three independent experiments; each color represents an independent experiment (***, P < 0.001; t test; error bars represent SD).

Article Snippet: UM-SCC-1 and UM-SCC-29 cells were transfected with empty vector (pCMV6) or DMBT1 plasmids (#RC215454; OriGene).

Techniques: Western Blot, Transfection, Over Expression, Transduction, Plasmid Preparation, Control, Incubation, Isolation, Reverse Transcription Polymerase Chain Reaction, Luciferase, Activity Assay

Overexpression of DMBT1 regulates mRNA stability and transcription of MMP9 and E-cadherin (E-cad). (A) Expression of E-cad and MMP9 in UM-SCC-1-pCMV6 and UM-SCC-1-DMBT1 cells ( n = 2). (B and C) UM-SCC-1-pCMV6 and UM-SCC-1-DMBT1 were incubated with actinomycin D (1 µg/ml), and RNA was isolated at different time points. MMP9 (B) and E-cad (C) transcripts were quantified by Q-RT-PCR. MMP9 and E-cad mRNA expression was normalized to GAPDH. Results are presented as percent of corresponding transcript at time 0. Values are mean ± SEM. (D and E) Normalized luciferase activity of MMP9 promoter (D) is reduced whereas activity of CDH1 promoter (E) is significantly higher in UM-SCC-1-DMBT1 compared with control UM-SCC-1-pCMV6 cells (*, P < 0.001; n = 2; t test; error bars represent SD). (F) Q-RT-PCR of UM-SCC-1-pCMV6, UM-SCC-1-DMBT1, UM-SCC-29-pCMV6, and UM-SCC-29-DMBT1. Primers for ZEB1, NRF2 (NFE2), β-catenin, VDR, YY1, c-Jun (AP1), c-Fos (AP1), SP1, EGR1, KLF4, NFATC1 (NFAT), HNF1 (HNF1A), STAT1, STAT3, and TP53 are listed in . Data are normalized to GAPDH. Relative expression of each transcription factor in cells overexpressing DMBT1 is shown as fold change with respect to control (pCMV6) cells (*, P < 0.05; t test; error bars represent SD).

Journal: The Journal of Experimental Medicine

Article Title: Squamous cell carcinoma subverts adjacent histologically normal epithelium to promote lateral invasion

doi: 10.1084/jem.20200944

Figure Lengend Snippet: Overexpression of DMBT1 regulates mRNA stability and transcription of MMP9 and E-cadherin (E-cad). (A) Expression of E-cad and MMP9 in UM-SCC-1-pCMV6 and UM-SCC-1-DMBT1 cells ( n = 2). (B and C) UM-SCC-1-pCMV6 and UM-SCC-1-DMBT1 were incubated with actinomycin D (1 µg/ml), and RNA was isolated at different time points. MMP9 (B) and E-cad (C) transcripts were quantified by Q-RT-PCR. MMP9 and E-cad mRNA expression was normalized to GAPDH. Results are presented as percent of corresponding transcript at time 0. Values are mean ± SEM. (D and E) Normalized luciferase activity of MMP9 promoter (D) is reduced whereas activity of CDH1 promoter (E) is significantly higher in UM-SCC-1-DMBT1 compared with control UM-SCC-1-pCMV6 cells (*, P < 0.001; n = 2; t test; error bars represent SD). (F) Q-RT-PCR of UM-SCC-1-pCMV6, UM-SCC-1-DMBT1, UM-SCC-29-pCMV6, and UM-SCC-29-DMBT1. Primers for ZEB1, NRF2 (NFE2), β-catenin, VDR, YY1, c-Jun (AP1), c-Fos (AP1), SP1, EGR1, KLF4, NFATC1 (NFAT), HNF1 (HNF1A), STAT1, STAT3, and TP53 are listed in . Data are normalized to GAPDH. Relative expression of each transcription factor in cells overexpressing DMBT1 is shown as fold change with respect to control (pCMV6) cells (*, P < 0.05; t test; error bars represent SD).

Article Snippet: UM-SCC-1 and UM-SCC-29 cells were transfected with empty vector (pCMV6) or DMBT1 plasmids (#RC215454; OriGene).

Techniques: Over Expression, Expressing, Incubation, Isolation, Reverse Transcription Polymerase Chain Reaction, Luciferase, Activity Assay, Control

ZEB1 down-regulates DMBT1 by binding promoter regions of MMP9 and CDH1 . (A) Venn diagram showing transcription factors with binding sites in the promoter regions of MMP9 and CDH1 , including ZEB1, NRF2 (NFE2), VDR, YY1, c-Jun (AP1), SP1, EGR1, KLF4, NFATC1 (NFAT), HNF1 (HNF1A), STAT1, STAT3, and TP53. (B and C) Q-RT-PCR of UM-SCC-1-pCMV6, UM-SCC-1-DMBT1, UM-SCC-29-pCMV6, and UM-SCC-29-DMBT1. Data are normalized to GAPDH. Relative expression of ZEB1 transcription factor in cells overexpressing DMBT1 is shown as fold change with respect to control (pCMV6) cells (***, P < 0.001; ****, P < 0.0001; t test; n = 2; error bars represent SD). (D and E) Q-RT-PCR was performed with ChIP-eluted DNA from UM-SCC-1-pCMV6, UM-SCC-1-DMBT1, UM-SCC-29-pCMV6, and UM-SCC-29-DMBT1 with validated primers specific to the ZEB1-binding site in the promoter region of MMP9 (D) and CDH1 (E; *, P < 0.05; t test; n = 2; error bars represent SD). (F) Immunoblot validation of DMBT1 down-regulation in lysates from UM-SCC-29-shVSVG and -shEZH2 transfected with nontarget siRNA (NT) or siDMBT1 (-1 and -2), as indicated. (G and H) Q-RT-PCR with ChIP-eluted DNA from UM-SCC-29-shVSVG and UM-SCC-29-shEZH2 (NT, siDMBT1-1, and siDMBT1-2) for standardized primers targeting the ZEB1-binding site in the promoter region of MMP9 (G) and CDH1 (H). Scatter plots show two independent experiments; each color represents an independent experiment (**, P < 0.01; ****, P < 0.0001; one-way ANOVA; error bars represent SD).

Journal: The Journal of Experimental Medicine

Article Title: Squamous cell carcinoma subverts adjacent histologically normal epithelium to promote lateral invasion

doi: 10.1084/jem.20200944

Figure Lengend Snippet: ZEB1 down-regulates DMBT1 by binding promoter regions of MMP9 and CDH1 . (A) Venn diagram showing transcription factors with binding sites in the promoter regions of MMP9 and CDH1 , including ZEB1, NRF2 (NFE2), VDR, YY1, c-Jun (AP1), SP1, EGR1, KLF4, NFATC1 (NFAT), HNF1 (HNF1A), STAT1, STAT3, and TP53. (B and C) Q-RT-PCR of UM-SCC-1-pCMV6, UM-SCC-1-DMBT1, UM-SCC-29-pCMV6, and UM-SCC-29-DMBT1. Data are normalized to GAPDH. Relative expression of ZEB1 transcription factor in cells overexpressing DMBT1 is shown as fold change with respect to control (pCMV6) cells (***, P < 0.001; ****, P < 0.0001; t test; n = 2; error bars represent SD). (D and E) Q-RT-PCR was performed with ChIP-eluted DNA from UM-SCC-1-pCMV6, UM-SCC-1-DMBT1, UM-SCC-29-pCMV6, and UM-SCC-29-DMBT1 with validated primers specific to the ZEB1-binding site in the promoter region of MMP9 (D) and CDH1 (E; *, P < 0.05; t test; n = 2; error bars represent SD). (F) Immunoblot validation of DMBT1 down-regulation in lysates from UM-SCC-29-shVSVG and -shEZH2 transfected with nontarget siRNA (NT) or siDMBT1 (-1 and -2), as indicated. (G and H) Q-RT-PCR with ChIP-eluted DNA from UM-SCC-29-shVSVG and UM-SCC-29-shEZH2 (NT, siDMBT1-1, and siDMBT1-2) for standardized primers targeting the ZEB1-binding site in the promoter region of MMP9 (G) and CDH1 (H). Scatter plots show two independent experiments; each color represents an independent experiment (**, P < 0.01; ****, P < 0.0001; one-way ANOVA; error bars represent SD).

Article Snippet: UM-SCC-1 and UM-SCC-29 cells were transfected with empty vector (pCMV6) or DMBT1 plasmids (#RC215454; OriGene).

Techniques: Binding Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Western Blot, Biomarker Discovery, Transfection