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Santa Cruz Biotechnology rabbit anti islet 1
Rabbit Anti Islet 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti islet 1/product/Santa Cruz Biotechnology
Average 85 stars, based on 1 article reviews
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rabbit anti islet 1 - by Bioz Stars, 2020-07
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CTG Assay:

Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes
Article Snippet: .. ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz). ..

Activated Clotting Time Assay:

Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes
Article Snippet: .. ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz). ..

Chromatin Immunoprecipitation:

Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes
Article Snippet: .. ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz). ..

Chloramphenicol Acetyltransferase Assay:

Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes
Article Snippet: .. ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz). ..

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  • 88
    Santa Cruz Biotechnology rabbit anti isl1
    ChIP-seq assays revealed Isl-Lhx3-hexamer-binding sites in a cholinergic gene battery. ( A ) Schematic representation of cholinergic neurotransmission system. Acly, ATP-citrate lyase; CoA, coenzyme A; ChAT, choline acetyltransferase; ACh, acetylcholine; VAChT, vesicular acetylcholine transporter; AChE, Acetylcholine esterase; CHT, high affinity choline transporter; AChRs, Acetylcholine receptors. ( B ) ChIP-seq tag profile of the genomic region surrounding a battery of cholinergic genes ChAT/VAChT , CHT , and Acly loci. Each cholinergic gene is indicated, and the blue arrows represent the direction of transcription. Mam cons., mammalian conservation. The ChIP-seq data was deposited in the GEO database (assession no. GSE50993) [20] . ( C ) Schematic representation of the location of the HxRE motifs in each of the 500 bp-long cholinergic gene peaks. The number shows the relative position within the peak (0, the center position of each peak). ( D ) In vivo ChIP assays in dissected E12.5 embryonic spinal cords to monitor the binding of the <t>Isl1-Lhx3-hexamer</t> to the cholinergic enhancers. Schematic representation of the ChAT gene is shown on the top. The arrows indicate two sets of primers detecting ChAT -enhancer ( ChAT -enh) and a negative control region lacking the Isl1-Lhx3-binding peak ( ChAT -neg). Isl1, Lhx3, and NLI were recruited to the cholinergic enhancers in embryonic spinal cords. Error bars indicate standard deviation.
    Rabbit Anti Isl1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti isl1/product/Santa Cruz Biotechnology
    Average 88 stars, based on 1 article reviews
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    91
    Santa Cruz Biotechnology rabbit anti pcna
    Hematoxylin and eosin (H E), and immunohistochemical (IHC) stain of glucagon, insulin, and proliferating cell nuclear antigen <t>(PCNA)</t> on islet before and after induction of type 2 diabetes mellitus in wild-type and uPA-/- mice and after treatment with uPA plasmids in uPA-/- mice. ( A ) The H E stain of islet showed no pathologic structural change in either wild-type or uPA-/- mice. The expression of glucagon on islet increased after induction both in wild-type and uPA-/- mice. Obviously, insulin particles of islet 30 days after induction (D30) increased in wild-type mice as compared with uPA-/- mice. Meanwhile, the PCNA expression of islet increased on D30 in wild-type mice. After treatment with uPA plasmids in uPA-/- mice, the glucagon, insulin, and PCNA expressions are similar to those on D30 in wild-type mice. ( B ) The quantitative intensity score of expression of glucagon, insulin, and PCNA on islets on induction day (D0), D3, and D30, after induction was assessed and scored by pathologists. The intensity scores of glucagon in uPA-/- mice were significantly higher than those in wild-type mice on D3. The intensity scores of insulin in wild-type mice were significantly higher than those in uPA-/- mice on D3 and D30. The intensity score of PCNA in wild-type mice on D30 was higher, but did not reach statistical significance. ** p
    Rabbit Anti Pcna, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti pcna/product/Santa Cruz Biotechnology
    Average 91 stars, based on 17 article reviews
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    85
    Santa Cruz Biotechnology rabbit polyclonal anti srebp 1
    Hematoxylin and eosin (H E), and immunohistochemical (IHC) stain of glucagon, insulin, and proliferating cell nuclear antigen <t>(PCNA)</t> on islet before and after induction of type 2 diabetes mellitus in wild-type and uPA-/- mice and after treatment with uPA plasmids in uPA-/- mice. ( A ) The H E stain of islet showed no pathologic structural change in either wild-type or uPA-/- mice. The expression of glucagon on islet increased after induction both in wild-type and uPA-/- mice. Obviously, insulin particles of islet 30 days after induction (D30) increased in wild-type mice as compared with uPA-/- mice. Meanwhile, the PCNA expression of islet increased on D30 in wild-type mice. After treatment with uPA plasmids in uPA-/- mice, the glucagon, insulin, and PCNA expressions are similar to those on D30 in wild-type mice. ( B ) The quantitative intensity score of expression of glucagon, insulin, and PCNA on islets on induction day (D0), D3, and D30, after induction was assessed and scored by pathologists. The intensity scores of glucagon in uPA-/- mice were significantly higher than those in wild-type mice on D3. The intensity scores of insulin in wild-type mice were significantly higher than those in uPA-/- mice on D3 and D30. The intensity score of PCNA in wild-type mice on D30 was higher, but did not reach statistical significance. ** p
    Rabbit Polyclonal Anti Srebp 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti srebp 1/product/Santa Cruz Biotechnology
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    90
    Santa Cruz Biotechnology rabbit anti isl1 210
    Hematoxylin and eosin (H E), and immunohistochemical (IHC) stain of glucagon, insulin, and proliferating cell nuclear antigen <t>(PCNA)</t> on islet before and after induction of type 2 diabetes mellitus in wild-type and uPA-/- mice and after treatment with uPA plasmids in uPA-/- mice. ( A ) The H E stain of islet showed no pathologic structural change in either wild-type or uPA-/- mice. The expression of glucagon on islet increased after induction both in wild-type and uPA-/- mice. Obviously, insulin particles of islet 30 days after induction (D30) increased in wild-type mice as compared with uPA-/- mice. Meanwhile, the PCNA expression of islet increased on D30 in wild-type mice. After treatment with uPA plasmids in uPA-/- mice, the glucagon, insulin, and PCNA expressions are similar to those on D30 in wild-type mice. ( B ) The quantitative intensity score of expression of glucagon, insulin, and PCNA on islets on induction day (D0), D3, and D30, after induction was assessed and scored by pathologists. The intensity scores of glucagon in uPA-/- mice were significantly higher than those in wild-type mice on D3. The intensity scores of insulin in wild-type mice were significantly higher than those in uPA-/- mice on D3 and D30. The intensity score of PCNA in wild-type mice on D30 was higher, but did not reach statistical significance. ** p
    Rabbit Anti Isl1 210, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ChIP-seq assays revealed Isl-Lhx3-hexamer-binding sites in a cholinergic gene battery. ( A ) Schematic representation of cholinergic neurotransmission system. Acly, ATP-citrate lyase; CoA, coenzyme A; ChAT, choline acetyltransferase; ACh, acetylcholine; VAChT, vesicular acetylcholine transporter; AChE, Acetylcholine esterase; CHT, high affinity choline transporter; AChRs, Acetylcholine receptors. ( B ) ChIP-seq tag profile of the genomic region surrounding a battery of cholinergic genes ChAT/VAChT , CHT , and Acly loci. Each cholinergic gene is indicated, and the blue arrows represent the direction of transcription. Mam cons., mammalian conservation. The ChIP-seq data was deposited in the GEO database (assession no. GSE50993) [20] . ( C ) Schematic representation of the location of the HxRE motifs in each of the 500 bp-long cholinergic gene peaks. The number shows the relative position within the peak (0, the center position of each peak). ( D ) In vivo ChIP assays in dissected E12.5 embryonic spinal cords to monitor the binding of the Isl1-Lhx3-hexamer to the cholinergic enhancers. Schematic representation of the ChAT gene is shown on the top. The arrows indicate two sets of primers detecting ChAT -enhancer ( ChAT -enh) and a negative control region lacking the Isl1-Lhx3-binding peak ( ChAT -neg). Isl1, Lhx3, and NLI were recruited to the cholinergic enhancers in embryonic spinal cords. Error bars indicate standard deviation.

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: ChIP-seq assays revealed Isl-Lhx3-hexamer-binding sites in a cholinergic gene battery. ( A ) Schematic representation of cholinergic neurotransmission system. Acly, ATP-citrate lyase; CoA, coenzyme A; ChAT, choline acetyltransferase; ACh, acetylcholine; VAChT, vesicular acetylcholine transporter; AChE, Acetylcholine esterase; CHT, high affinity choline transporter; AChRs, Acetylcholine receptors. ( B ) ChIP-seq tag profile of the genomic region surrounding a battery of cholinergic genes ChAT/VAChT , CHT , and Acly loci. Each cholinergic gene is indicated, and the blue arrows represent the direction of transcription. Mam cons., mammalian conservation. The ChIP-seq data was deposited in the GEO database (assession no. GSE50993) [20] . ( C ) Schematic representation of the location of the HxRE motifs in each of the 500 bp-long cholinergic gene peaks. The number shows the relative position within the peak (0, the center position of each peak). ( D ) In vivo ChIP assays in dissected E12.5 embryonic spinal cords to monitor the binding of the Isl1-Lhx3-hexamer to the cholinergic enhancers. Schematic representation of the ChAT gene is shown on the top. The arrows indicate two sets of primers detecting ChAT -enhancer ( ChAT -enh) and a negative control region lacking the Isl1-Lhx3-binding peak ( ChAT -neg). Isl1, Lhx3, and NLI were recruited to the cholinergic enhancers in embryonic spinal cords. Error bars indicate standard deviation.

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Chromatin Immunoprecipitation, Binding Assay, In Vivo, Negative Control, Standard Deviation

    The formation of the Isl1-Lhx8-hexamer complex. ( A ) Schematic representation of the Isl1-Lhx8-hexamer consisting of Isl1, Lhx8 and NLI. The model depicts that the Isl1-Lhx8-complex regulates the cholinergic genes via binding to HxREs. ( B ) In vitro GST-pull down assays. Lhx8 and Lhx3 bind to both Isl1 and NLI with high affinity, whereas Lhx1 binds to only NLI, but not to Isl1. ( C ) GST-pull down assays in HEK293 cells transfected with Flag- and GST-tagged constructs as indicated above. Lhx8 interacts with both Isl1 and NLI in cells. ( D ) CoIP assays in HEK293 cells transfected with Flag-Lhx8 and HA-tagged NLI DD -Isl1 ΔLIM . Lhx8 interact with NLI DD -Isl1 ΔLIM , forming the FCN-hexamer-mimicking complex. ( E, F ) The SELEX methods revealed the high affinity binding sites for Isl1-Lhx8 fusion (E-value, 2.5e-79) and the mixture of Isl1 and Lhx8 (E-value, 2.8e-65). The bottom sequence logo shows reverse complementary sequences of the upper logo.

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: The formation of the Isl1-Lhx8-hexamer complex. ( A ) Schematic representation of the Isl1-Lhx8-hexamer consisting of Isl1, Lhx8 and NLI. The model depicts that the Isl1-Lhx8-complex regulates the cholinergic genes via binding to HxREs. ( B ) In vitro GST-pull down assays. Lhx8 and Lhx3 bind to both Isl1 and NLI with high affinity, whereas Lhx1 binds to only NLI, but not to Isl1. ( C ) GST-pull down assays in HEK293 cells transfected with Flag- and GST-tagged constructs as indicated above. Lhx8 interacts with both Isl1 and NLI in cells. ( D ) CoIP assays in HEK293 cells transfected with Flag-Lhx8 and HA-tagged NLI DD -Isl1 ΔLIM . Lhx8 interact with NLI DD -Isl1 ΔLIM , forming the FCN-hexamer-mimicking complex. ( E, F ) The SELEX methods revealed the high affinity binding sites for Isl1-Lhx8 fusion (E-value, 2.5e-79) and the mixture of Isl1 and Lhx8 (E-value, 2.8e-65). The bottom sequence logo shows reverse complementary sequences of the upper logo.

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Binding Assay, In Vitro, Transfection, Construct, Co-Immunoprecipitation Assay, Sequencing

    Isl1 is required for the formation of striatal cholinergic interneurons in the developing forebrain. Immunohistochemical analyses on the CPu (A, B) and BMC (C, D) of E16.5 Isl1 f/f ;Nkx2.1Cre and littermate control embryos. VAChT + cholinergic interneurons in the CPu failed to form in the MGE-specific Isl1 -null embryos. (E, F) Quantification of the number of Lhx8 + VAChT + (E) or Nkx2.1 + VAChT + cells in the CPu of E16.5 control and Isl1 mutant embryos. Histogram shows average ± standard deviation. ** p

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: Isl1 is required for the formation of striatal cholinergic interneurons in the developing forebrain. Immunohistochemical analyses on the CPu (A, B) and BMC (C, D) of E16.5 Isl1 f/f ;Nkx2.1Cre and littermate control embryos. VAChT + cholinergic interneurons in the CPu failed to form in the MGE-specific Isl1 -null embryos. (E, F) Quantification of the number of Lhx8 + VAChT + (E) or Nkx2.1 + VAChT + cells in the CPu of E16.5 control and Isl1 mutant embryos. Histogram shows average ± standard deviation. ** p

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Immunohistochemistry, Mutagenesis, Standard Deviation

    Isl1-Lhx8 induces a cholinergic fate in ESC-derived neurons. ( A, B ) In Isl1-Lhx8-ESCs, the expression of Flag-tagged Isl1-Lhx8 was induced by Dox, as detected by western blotting assays with α-Flag antibodies (A) and immunohistochemistry assays with α-Isl1 and α-Lhx8 antibodies (B). ( C, D ) Quantitative RT-PCR analyses in Isl1-Lhx8-ESCs when cultured as a monolayer. Cholinergic genes, but not the MN gene Hb9, were induced by Isl1-Lhx8. ( E–G ) Cell differentiation analyses in floating EBs derived from Isl1-Lhx8-ESCs, cultured with or without Dox, which triggers expression of Isl1-Lhx8. Immunohistochemical analyses show that Isl-Lhx8 expression induces differentiation of VAChT + TuJ + c holinergic neurons (E, F). Quantitative RT-PCR analyses show that cholinergic pathway genes, but not MN genes Hb9 and Isl2, were induced by Isl1-Lhx8 (G). ( H ) Quantitative RT-PCR analyses of Hb9 expression in Isl1-Lhx3-ESCs. Hb9 was induced by Dox treatment, which induces the expression of Isl1-Lhx3 in Isl1-Lhx3-ESCs, when cultured in either monolayer (M) or spinal neuronal differentiation (SN) conditions. Error bars represent the standard deviation in all graphs (C, D, G, H).

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: Isl1-Lhx8 induces a cholinergic fate in ESC-derived neurons. ( A, B ) In Isl1-Lhx8-ESCs, the expression of Flag-tagged Isl1-Lhx8 was induced by Dox, as detected by western blotting assays with α-Flag antibodies (A) and immunohistochemistry assays with α-Isl1 and α-Lhx8 antibodies (B). ( C, D ) Quantitative RT-PCR analyses in Isl1-Lhx8-ESCs when cultured as a monolayer. Cholinergic genes, but not the MN gene Hb9, were induced by Isl1-Lhx8. ( E–G ) Cell differentiation analyses in floating EBs derived from Isl1-Lhx8-ESCs, cultured with or without Dox, which triggers expression of Isl1-Lhx8. Immunohistochemical analyses show that Isl-Lhx8 expression induces differentiation of VAChT + TuJ + c holinergic neurons (E, F). Quantitative RT-PCR analyses show that cholinergic pathway genes, but not MN genes Hb9 and Isl2, were induced by Isl1-Lhx8 (G). ( H ) Quantitative RT-PCR analyses of Hb9 expression in Isl1-Lhx3-ESCs. Hb9 was induced by Dox treatment, which induces the expression of Isl1-Lhx3 in Isl1-Lhx3-ESCs, when cultured in either monolayer (M) or spinal neuronal differentiation (SN) conditions. Error bars represent the standard deviation in all graphs (C, D, G, H).

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Derivative Assay, Expressing, Western Blot, Immunohistochemistry, Quantitative RT-PCR, Cell Culture, Cell Differentiation, Standard Deviation

    The cholinergic enhancers are activated by the Isl1-Lhx3-hexamer in the developing spinal cord. ( A–E ) Luciferase reporter assays in mouse embryonic P19 cells using Acly -enh1-wt:LUC or Acly -enh1-mt:LUC, in which HxRE motifs are mutated (A), ChAT -enh:LUC (B), CHT -enh:LUC (C), Acly -HxRE:LUC (D), and ChAT -HxRE:LUC (E) reporters with expression vectors as indicated below each graph. The co-expression of Lhx3 wild-type and Isl1 wild-type, indicated as w or +, strongly activated the reporters linked to the cholinergic enhancers, but not the LUC vector alone or Acly -enh1-mt:LUC. Lhx3-N211S and Isl1-N230S, the DNA-binding defective missense mutants of Lhx3 or Isl1 that are indicated as m, failed to activate Acly -enh1-wt:LUC (A). (A–E) Error bars represent the standard deviation in all graphs. Error bars indicate standard deviation. ( F–H ) GFP reporter activity was monitored in chick embryos electroporated with Acly -enh1:GFP (F), Acly -enh1-HxRE-mt:GFP (G), and Acly -HxRE:GFP (H) reporters with either LacZ or Isl1 plus Lhx3 as indicated above. Acly -enh1 and Acly -HxRE drove MN-specific GFP expression, and were ectopically activated by co-expression of Isl1 and Lhx3 in the dorsal spinal cord (F, H). Acly -enh1-HxRE-mt:GFP did not display GFP expression in MNs and failed to respond to the co-electroporated Isl1 and Lhx3 (G), indicating that the HxRE motif is required for the MN-specific enhancer activity of Acly -enh1. + indicates the electroporated side. The areas of ectopic Hb9 + MNs, induced by co-expression of Isl1 and Lhx3, are marked by brackets.

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: The cholinergic enhancers are activated by the Isl1-Lhx3-hexamer in the developing spinal cord. ( A–E ) Luciferase reporter assays in mouse embryonic P19 cells using Acly -enh1-wt:LUC or Acly -enh1-mt:LUC, in which HxRE motifs are mutated (A), ChAT -enh:LUC (B), CHT -enh:LUC (C), Acly -HxRE:LUC (D), and ChAT -HxRE:LUC (E) reporters with expression vectors as indicated below each graph. The co-expression of Lhx3 wild-type and Isl1 wild-type, indicated as w or +, strongly activated the reporters linked to the cholinergic enhancers, but not the LUC vector alone or Acly -enh1-mt:LUC. Lhx3-N211S and Isl1-N230S, the DNA-binding defective missense mutants of Lhx3 or Isl1 that are indicated as m, failed to activate Acly -enh1-wt:LUC (A). (A–E) Error bars represent the standard deviation in all graphs. Error bars indicate standard deviation. ( F–H ) GFP reporter activity was monitored in chick embryos electroporated with Acly -enh1:GFP (F), Acly -enh1-HxRE-mt:GFP (G), and Acly -HxRE:GFP (H) reporters with either LacZ or Isl1 plus Lhx3 as indicated above. Acly -enh1 and Acly -HxRE drove MN-specific GFP expression, and were ectopically activated by co-expression of Isl1 and Lhx3 in the dorsal spinal cord (F, H). Acly -enh1-HxRE-mt:GFP did not display GFP expression in MNs and failed to respond to the co-electroporated Isl1 and Lhx3 (G), indicating that the HxRE motif is required for the MN-specific enhancer activity of Acly -enh1. + indicates the electroporated side. The areas of ectopic Hb9 + MNs, induced by co-expression of Isl1 and Lhx3, are marked by brackets.

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Luciferase, Expressing, Plasmid Preparation, Binding Assay, Standard Deviation, Activity Assay

    The Isl1-Lhx8-hexamer activates the cholinergic genes. ( A ) ChIP assays with IgG, α-Isl1, α-Lhx8, and α-NLI antibodies in dissected E15.5 embryonic forebrains. The location of two sets of primers in the ChAT gene is indicated (arrows). The FCN-hexamer is recruited to the cholinergic enhancers. ( B, C ) Luciferase reporter assays in P19 cells using Acly -HxRE:LUC (B), and ChAT -HxRE:LUC (C) reporters with vectors indicated below each graph. The co-expression of Lhx8 and Isl1 strongly activated these reporters. Error bars represent the standard deviation in all graphs (A–C). ( D ) Schematic representation of ex vivo electroporation of the ventral forebrain. Sections containing the appropriate regions of the ventral forebrain were focally injected with combinations of plasmids and subjected to slice electroporation, followed by slice culture. The area of transfection was indicated in green. Due to the electroporation process with slices, part of cortex was also transfected with plasmids. ( E–H ) Activation of Acly -HxRE:GFP reporter in the ventral forebrains electroporated with constructs, LacZ (E), Lhx8 (F), Isl1 (G), and Isl1 and Lhx8 (H). The co-expression of Isl1 and Lhx8 strongly activated Acly -HxRE in the forebrain.

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: The Isl1-Lhx8-hexamer activates the cholinergic genes. ( A ) ChIP assays with IgG, α-Isl1, α-Lhx8, and α-NLI antibodies in dissected E15.5 embryonic forebrains. The location of two sets of primers in the ChAT gene is indicated (arrows). The FCN-hexamer is recruited to the cholinergic enhancers. ( B, C ) Luciferase reporter assays in P19 cells using Acly -HxRE:LUC (B), and ChAT -HxRE:LUC (C) reporters with vectors indicated below each graph. The co-expression of Lhx8 and Isl1 strongly activated these reporters. Error bars represent the standard deviation in all graphs (A–C). ( D ) Schematic representation of ex vivo electroporation of the ventral forebrain. Sections containing the appropriate regions of the ventral forebrain were focally injected with combinations of plasmids and subjected to slice electroporation, followed by slice culture. The area of transfection was indicated in green. Due to the electroporation process with slices, part of cortex was also transfected with plasmids. ( E–H ) Activation of Acly -HxRE:GFP reporter in the ventral forebrains electroporated with constructs, LacZ (E), Lhx8 (F), Isl1 (G), and Isl1 and Lhx8 (H). The co-expression of Isl1 and Lhx8 strongly activated Acly -HxRE in the forebrain.

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Chromatin Immunoprecipitation, Luciferase, Expressing, Standard Deviation, Ex Vivo, Electroporation, Injection, Transfection, Activation Assay, Construct

    Isl1-Lhx8 induces the expression of cholinergic gene battery in the forebrain, but not in the spinal cord. ( A ) Schematic representation of in utero electroporation of the cortex, followed by quantitative RT-PCR (qRT-PCR) analyses. E13.5 brains were subjected to electroporation after each combination of constructs was injected into the lateral ventricle. The GFP + region of electroporated (+) cortex and the comparable region of unelectroporated (−) cortex were micro-dissected and analyzed for gene expression. ( B ) Expression analyses of the cholinergic pathway genes, ChAT, VAChT and CHT , in mouse cortices electroporated with constructs, as indicated by color bars. Y-axis indicates the relative expression levels of each cholinergic gene on the electropoated side over the control side. The expression of Isl1-Lhx8 led to upregulation of the cholinergic genes in the cortex, whereas that of Isl1-Lhx3 failed to induce cholinergic genes in this context. Error bars indicate standard deviation. ** p

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: Isl1-Lhx8 induces the expression of cholinergic gene battery in the forebrain, but not in the spinal cord. ( A ) Schematic representation of in utero electroporation of the cortex, followed by quantitative RT-PCR (qRT-PCR) analyses. E13.5 brains were subjected to electroporation after each combination of constructs was injected into the lateral ventricle. The GFP + region of electroporated (+) cortex and the comparable region of unelectroporated (−) cortex were micro-dissected and analyzed for gene expression. ( B ) Expression analyses of the cholinergic pathway genes, ChAT, VAChT and CHT , in mouse cortices electroporated with constructs, as indicated by color bars. Y-axis indicates the relative expression levels of each cholinergic gene on the electropoated side over the control side. The expression of Isl1-Lhx8 led to upregulation of the cholinergic genes in the cortex, whereas that of Isl1-Lhx3 failed to induce cholinergic genes in this context. Error bars indicate standard deviation. ** p

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Expressing, In Utero, Electroporation, Quantitative RT-PCR, Construct, Injection, Standard Deviation

    Isl1 is co-expressed with Nkx2.1 and Lhx8 in the CPu and BMC and is important for the formation of Nkx2.1/Lhx8-expressing striatal interneurons. ( A ) Schematic representation of the coronal section of E16.5 forebrain. CPu, Caudate-putamen; BMC, basal meganocellular complex. ( B–I ) Immunohistochemical analyses on the CPu and BMC of E16.5 Isl1 f/f ;Nkx2.1Cre and littermate control embryos. Isl1 is co-expressed with Nkx2.1 and Lhx8 in subsets of neurons in the CPu and the BMC (B, C, F, G). The dotted circles depict Isl1/Nkx2.1-double positive cells (D, F). In Isl1 f/f ;Nkx2.1Cre embryos, the number of Nkx2.1 or Lhx8-expressing interneurons in the CPu is reduced (B–E), and Isl1 + cells in the BMC drastically decreased (F–I). ( J, K ) Quantification of the number of Lhx8- and Nkx2.1-expressing cells in the CPu of E16.5 control and Isl1 mutant embryos. Histogram shows average ± standard deviation. ** p

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: Isl1 is co-expressed with Nkx2.1 and Lhx8 in the CPu and BMC and is important for the formation of Nkx2.1/Lhx8-expressing striatal interneurons. ( A ) Schematic representation of the coronal section of E16.5 forebrain. CPu, Caudate-putamen; BMC, basal meganocellular complex. ( B–I ) Immunohistochemical analyses on the CPu and BMC of E16.5 Isl1 f/f ;Nkx2.1Cre and littermate control embryos. Isl1 is co-expressed with Nkx2.1 and Lhx8 in subsets of neurons in the CPu and the BMC (B, C, F, G). The dotted circles depict Isl1/Nkx2.1-double positive cells (D, F). In Isl1 f/f ;Nkx2.1Cre embryos, the number of Nkx2.1 or Lhx8-expressing interneurons in the CPu is reduced (B–E), and Isl1 + cells in the BMC drastically decreased (F–I). ( J, K ) Quantification of the number of Lhx8- and Nkx2.1-expressing cells in the CPu of E16.5 control and Isl1 mutant embryos. Histogram shows average ± standard deviation. ** p

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Expressing, Immunohistochemistry, Mutagenesis, Standard Deviation

    The Isl1-Lhx3-hexamer plays a crucial role in inducing the expression of cholinergic pathway genes in the developing spinal MNs. ( A ) Expression analyses of the cholinergic pathway genes in chick embryos electroporated with Isl1 and Lhx3 using in situ hybridization. The co-electroporation of Isl1 and Lhx3 triggered the ectopic expression of cholinergic genes in the dorsal spinal cord, as marked by brackets. + indicates the electroporated side. ( B ) Expression analyses of the cholinergic pathway genes using either immunohistochemistry or in situ hybridization on the spinal cord of E12.5 Isl1 f/f ;nestinCre and littermate control embryos. The ventral quadrant spinal cord is shown. The cholinergic genes are markedly downregulated in Isl1 f/f ;nestinCre mice. The remaining VAChT expression is correlated with the residual Isl1 expression in Isl1 f/f ;nestinCre mice, as determined by immunostaining assays.

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: The Isl1-Lhx3-hexamer plays a crucial role in inducing the expression of cholinergic pathway genes in the developing spinal MNs. ( A ) Expression analyses of the cholinergic pathway genes in chick embryos electroporated with Isl1 and Lhx3 using in situ hybridization. The co-electroporation of Isl1 and Lhx3 triggered the ectopic expression of cholinergic genes in the dorsal spinal cord, as marked by brackets. + indicates the electroporated side. ( B ) Expression analyses of the cholinergic pathway genes using either immunohistochemistry or in situ hybridization on the spinal cord of E12.5 Isl1 f/f ;nestinCre and littermate control embryos. The ventral quadrant spinal cord is shown. The cholinergic genes are markedly downregulated in Isl1 f/f ;nestinCre mice. The remaining VAChT expression is correlated with the residual Isl1 expression in Isl1 f/f ;nestinCre mice, as determined by immunostaining assays.

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Expressing, In Situ Hybridization, Electroporation, Immunohistochemistry, Mouse Assay, Immunostaining

    Isl1-Lhx8-hexamer and Isl1-Lhx3-hexamer complexes establish a cholinergic neuronal identity in FCNs and spinal MNs, respectively, by directly upregulating cholinergic gene battery. During CNS development, the cholinergic genes recruit the Isl1-Lhx8-hexamer in the forebrain and the Isl1-Lhx3-hexamer in spinal cord via hexamer-response elements. This recruitment leads to concerted induction of the cholinergic genes, therefore enabling MNs and FCNs to acquire the cholinergic neuronal identity. The Isl1-Lhx8-hexamer and Isl1-Lhx3-hexamer likely induce unique sets of target genes in FCNs and MNs. These hexamers may cooperate with other transcription factors (TFs) in establishing cell type-specific gene expression patterns.

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: Isl1-Lhx8-hexamer and Isl1-Lhx3-hexamer complexes establish a cholinergic neuronal identity in FCNs and spinal MNs, respectively, by directly upregulating cholinergic gene battery. During CNS development, the cholinergic genes recruit the Isl1-Lhx8-hexamer in the forebrain and the Isl1-Lhx3-hexamer in spinal cord via hexamer-response elements. This recruitment leads to concerted induction of the cholinergic genes, therefore enabling MNs and FCNs to acquire the cholinergic neuronal identity. The Isl1-Lhx8-hexamer and Isl1-Lhx3-hexamer likely induce unique sets of target genes in FCNs and MNs. These hexamers may cooperate with other transcription factors (TFs) in establishing cell type-specific gene expression patterns.

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Expressing

    Co-expression of Isl1, Lhx8 and NLI in the developing ventral forebrain. ( A ) Schematic representation of the coronal section of E12.5 forebrain. The MGE produces striatal cholinergic interneurons in the CPu and cholinergic projection neurons in the BMC, which take different migratory paths. NCx, neocortex; MGE, medial ganaglionic eminence; LGE, lateral ganglionic eminence; CPu, Caudate-putamen; BMC, basal meganocellular complex. ( B–E ) Immunohistochemical analyses of expression of Nkx2.1, Isl1, Lhx8, and NLI on coronal sections of E12.5 mouse forebrains. Isl1 is co-expressed with Lhx8 and NLI in the mantle zone of the MGE (yellow asterisk) and LGE (red asterisk).

    Journal: PLoS Genetics

    Article Title: Isl1 Directly Controls a Cholinergic Neuronal Identity in the Developing Forebrain and Spinal Cord by Forming Cell Type-Specific Complexes

    doi: 10.1371/journal.pgen.1004280

    Figure Lengend Snippet: Co-expression of Isl1, Lhx8 and NLI in the developing ventral forebrain. ( A ) Schematic representation of the coronal section of E12.5 forebrain. The MGE produces striatal cholinergic interneurons in the CPu and cholinergic projection neurons in the BMC, which take different migratory paths. NCx, neocortex; MGE, medial ganaglionic eminence; LGE, lateral ganglionic eminence; CPu, Caudate-putamen; BMC, basal meganocellular complex. ( B–E ) Immunohistochemical analyses of expression of Nkx2.1, Isl1, Lhx8, and NLI on coronal sections of E12.5 mouse forebrains. Isl1 is co-expressed with Lhx8 and NLI in the mantle zone of the MGE (yellow asterisk) and LGE (red asterisk).

    Article Snippet: ChAT -enhancer forward 5′-TAC TAA TTG GAT TAA TTG ATT TGC reverse 5′-GGG AAT TAA TAA CTT AGA ATT TGA ChAT -negative forward 5′- CTG TGG CTC ATA ACG CTC ATT TTG reverse 5′- AGT TTG TGG TGG GCC GAG ATG GCA Acly -enh1 forward 5′- TGA TAG CAC ACT ACT TTG CTC TGG reverse 5′-CAG TGA CGC ACG GCG AGC GGG AAG CHT -enhancer forward 5′-TGA GCA GCC TAT GCC ACA AGG ACA reverse 5′- CAT TAG GAG AGC TTG TTC CAG TGA The following antibodies were used for ChIP-PCR; mouse/rabbit IgG (Santa Cruz), rabbit anti-Isl1 , rabbit anti-Lhx3 , rabbit anti-NLI , and goat anti-Lhx8 (sc-22216, Santa Cruz).

    Techniques: Expressing, Immunohistochemistry

    Hematoxylin and eosin (H E), and immunohistochemical (IHC) stain of glucagon, insulin, and proliferating cell nuclear antigen (PCNA) on islet before and after induction of type 2 diabetes mellitus in wild-type and uPA-/- mice and after treatment with uPA plasmids in uPA-/- mice. ( A ) The H E stain of islet showed no pathologic structural change in either wild-type or uPA-/- mice. The expression of glucagon on islet increased after induction both in wild-type and uPA-/- mice. Obviously, insulin particles of islet 30 days after induction (D30) increased in wild-type mice as compared with uPA-/- mice. Meanwhile, the PCNA expression of islet increased on D30 in wild-type mice. After treatment with uPA plasmids in uPA-/- mice, the glucagon, insulin, and PCNA expressions are similar to those on D30 in wild-type mice. ( B ) The quantitative intensity score of expression of glucagon, insulin, and PCNA on islets on induction day (D0), D3, and D30, after induction was assessed and scored by pathologists. The intensity scores of glucagon in uPA-/- mice were significantly higher than those in wild-type mice on D3. The intensity scores of insulin in wild-type mice were significantly higher than those in uPA-/- mice on D3 and D30. The intensity score of PCNA in wild-type mice on D30 was higher, but did not reach statistical significance. ** p

    Journal: Molecules

    Article Title: Deficiency of Urokinase Plasminogen Activator May Impair β Cells Regeneration and Insulin Secretion in Type 2 Diabetes Mellitus

    doi: 10.3390/molecules24234208

    Figure Lengend Snippet: Hematoxylin and eosin (H E), and immunohistochemical (IHC) stain of glucagon, insulin, and proliferating cell nuclear antigen (PCNA) on islet before and after induction of type 2 diabetes mellitus in wild-type and uPA-/- mice and after treatment with uPA plasmids in uPA-/- mice. ( A ) The H E stain of islet showed no pathologic structural change in either wild-type or uPA-/- mice. The expression of glucagon on islet increased after induction both in wild-type and uPA-/- mice. Obviously, insulin particles of islet 30 days after induction (D30) increased in wild-type mice as compared with uPA-/- mice. Meanwhile, the PCNA expression of islet increased on D30 in wild-type mice. After treatment with uPA plasmids in uPA-/- mice, the glucagon, insulin, and PCNA expressions are similar to those on D30 in wild-type mice. ( B ) The quantitative intensity score of expression of glucagon, insulin, and PCNA on islets on induction day (D0), D3, and D30, after induction was assessed and scored by pathologists. The intensity scores of glucagon in uPA-/- mice were significantly higher than those in wild-type mice on D3. The intensity scores of insulin in wild-type mice were significantly higher than those in uPA-/- mice on D3 and D30. The intensity score of PCNA in wild-type mice on D30 was higher, but did not reach statistical significance. ** p

    Article Snippet: For IHC staining of PCNA in islet, the rabbit anti-PCNA (1:200; Santa Cruz Biotechnology) was the primary antibody and the goat anti-rabbit IgG-HRP (1:200; Invitrogen, Life Technologies) was the secondary antibody.

    Techniques: Immunohistochemistry, Staining, Mouse Assay, Expressing