anti isl1 islet 1  (Abcam)

 
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    Name:
    Anti Islet 1 antibody EP4182
    Description:

    Catalog Number:
    ab109517
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    Structured Review

    Abcam anti isl1 islet 1

    https://www.bioz.com/result/anti isl1 islet 1/product/Abcam
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti isl1 islet 1 - by Bioz Stars, 2020-07
    94/100 stars

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    Western Blot:

    Article Title: Embryonic maturation of epidermal Merkel cells is controlled by a redundant transcription factor network
    Article Snippet: .. Western blot analysis was performed using anti-Isl1 (Abcam, ab109517; 1:1000), anti-Sox2 (Stemgent, 09-0024; 1:1000) and anti-Histone H3 (Abcam, ab1791; 1:10,000). ..

    Incubation:

    Article Title: ISL1 and JMJD3 synergistically control cardiac differentiation of embryonic stem cells
    Article Snippet: .. Specifically, 1 mg NE was incubated antibodies against 2 μg Isl1 (ab109517, Abcam) or 5 μg Flag (F3165, Sigma) in a volume of 400 μl Buffer C, supplemented with 200 μl Buffer BN (20 mM Tris–HCl, pH 8.0, 100 mM KCl, 0.2 mM EDTA, 20% glycerol, 0.5 mg/ml BSA, 0.1% NP-40, 0.5 mM PMSF, 1 μg/ml Pepstatin A, 1 μg/ml Leupeptin, 1 μg/ml Aprotinin). .. After overnight incubation at 4°C, 30 μl of protein A/G (1:1) beads were added for incubation at 4°C for 2 h. Beads were then washed with Buffer BN for 4 times and boiled in 2 × SDS sample buffer.

    Immunoprecipitation:

    Article Title: Collaborative ISL1/GATA3 interaction in controlling neuroblastoma oncogenic pathways overlapping with but distinct from MYCN
    Article Snippet: .. ISL1-bound DNA was immunoprecipitated by anti-ISL1 antibody (ab109517, Abcam, MA, USA). .. After reverse cross-linking, the DNA was purified using the Qiagen PCR purification kit (Qiagen).

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  • 90
    Abcam mouse anti isl1
    Figure 6. Structure of cardiomyocyte colonies grown in the primary culture of rat neonatal myocardial cells. ( A–C ) Different stages of development of the colonies stemming from <t>Isl1</t> + CSCs. ( A ) Cell division, DIV 2. Isl1 + (FITC, green), GATA-4 (phycoerythrin, red). ( B ) Colony consisting of approximately 8 cells, DIV 11. Isl1 + (FITC, green), actin (rhodamine-phalloidin, red). ( C ) Large Isl1 + colony, DIV 11. Isl1 + (FITC, green), actin (rhodamine-phalloidin, red). ( D ) The optical sections of colonies formed by Isl1 + , c-kit + , and Sca1 + CSCs on the 11th DIV. Isl1 + CSCs (Alexa 405, blue), Z = 12. c-kit + CSCs (FITC, green), Z = 12. Sca1 + CSCs (Alexa 405, blue), Z = 11. Actin was stained using rhodamine-phalloidin (red). ( E ) Differentiation of c-kit + CSCs inside the colony on the 13th DIV. Overlaid optical section of transmitted light and fluorescent images in 2 emitting wavelengths: 488 nm (FITC) and 543 nm (Alexa) in the bottom (Z = 5), in the middle (Z = 10), and the top (Z = 20) parts of the colony. c-kit + expression was revealed by FITC-conjugated antibodies (green), and α-sarcomeric actinin was revealed by Alexa-conjugated antibodies (red). Confocal microscope, Leica TCS SP5 (Germany), objective ×63, oil.
    Mouse Anti Isl1, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti isl1/product/Abcam
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    mouse anti isl1 - by Bioz Stars, 2020-07
    90/100 stars
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    94
    Abcam rabbit anti isl1
    <t>ISL1</t> directly regulates a number of genes required for normal pacemaker function in mice and human.
    Rabbit Anti Isl1, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti isl1/product/Abcam
    Average 94 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    rabbit anti isl1 - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    Image Search Results


    Figure 6. Structure of cardiomyocyte colonies grown in the primary culture of rat neonatal myocardial cells. ( A–C ) Different stages of development of the colonies stemming from Isl1 + CSCs. ( A ) Cell division, DIV 2. Isl1 + (FITC, green), GATA-4 (phycoerythrin, red). ( B ) Colony consisting of approximately 8 cells, DIV 11. Isl1 + (FITC, green), actin (rhodamine-phalloidin, red). ( C ) Large Isl1 + colony, DIV 11. Isl1 + (FITC, green), actin (rhodamine-phalloidin, red). ( D ) The optical sections of colonies formed by Isl1 + , c-kit + , and Sca1 + CSCs on the 11th DIV. Isl1 + CSCs (Alexa 405, blue), Z = 12. c-kit + CSCs (FITC, green), Z = 12. Sca1 + CSCs (Alexa 405, blue), Z = 11. Actin was stained using rhodamine-phalloidin (red). ( E ) Differentiation of c-kit + CSCs inside the colony on the 13th DIV. Overlaid optical section of transmitted light and fluorescent images in 2 emitting wavelengths: 488 nm (FITC) and 543 nm (Alexa) in the bottom (Z = 5), in the middle (Z = 10), and the top (Z = 20) parts of the colony. c-kit + expression was revealed by FITC-conjugated antibodies (green), and α-sarcomeric actinin was revealed by Alexa-conjugated antibodies (red). Confocal microscope, Leica TCS SP5 (Germany), objective ×63, oil.

    Journal: Cell Cycle

    Article Title: Characterization of contracting cardiomyocyte colonies in the primary culture of neonatal rat myocardial cells

    doi: 10.4161/cc.27768

    Figure Lengend Snippet: Figure 6. Structure of cardiomyocyte colonies grown in the primary culture of rat neonatal myocardial cells. ( A–C ) Different stages of development of the colonies stemming from Isl1 + CSCs. ( A ) Cell division, DIV 2. Isl1 + (FITC, green), GATA-4 (phycoerythrin, red). ( B ) Colony consisting of approximately 8 cells, DIV 11. Isl1 + (FITC, green), actin (rhodamine-phalloidin, red). ( C ) Large Isl1 + colony, DIV 11. Isl1 + (FITC, green), actin (rhodamine-phalloidin, red). ( D ) The optical sections of colonies formed by Isl1 + , c-kit + , and Sca1 + CSCs on the 11th DIV. Isl1 + CSCs (Alexa 405, blue), Z = 12. c-kit + CSCs (FITC, green), Z = 12. Sca1 + CSCs (Alexa 405, blue), Z = 11. Actin was stained using rhodamine-phalloidin (red). ( E ) Differentiation of c-kit + CSCs inside the colony on the 13th DIV. Overlaid optical section of transmitted light and fluorescent images in 2 emitting wavelengths: 488 nm (FITC) and 543 nm (Alexa) in the bottom (Z = 5), in the middle (Z = 10), and the top (Z = 20) parts of the colony. c-kit + expression was revealed by FITC-conjugated antibodies (green), and α-sarcomeric actinin was revealed by Alexa-conjugated antibodies (red). Confocal microscope, Leica TCS SP5 (Germany), objective ×63, oil.

    Article Snippet: In the second series, the primary mouse anti-Isl1 (Abcam) and anti-Sca1 (Abcam) monoclonal antibodies were preliminary conjugated with Alexa 405 according to Zenon technology (Invitrogen) and then used for immunostaining at a 1:100 dilution.

    Techniques: Staining, Expressing, Microscopy

    Figure 7. Differentiation of Isl1 + and c-kit + CSCs inside the colonies on the 13th DIV. The optical sections of colonies on 2 levels: ( A ) Isl1 + middle (Z = 14) and ( B ) bottom (Z = 0). ( C ) c-kit + top (Z = 10) and ( D ) bottom (Z = 0). Confocal microscope, LEICA TCS SL, objective ×63, oil.

    Journal: Cell Cycle

    Article Title: Characterization of contracting cardiomyocyte colonies in the primary culture of neonatal rat myocardial cells

    doi: 10.4161/cc.27768

    Figure Lengend Snippet: Figure 7. Differentiation of Isl1 + and c-kit + CSCs inside the colonies on the 13th DIV. The optical sections of colonies on 2 levels: ( A ) Isl1 + middle (Z = 14) and ( B ) bottom (Z = 0). ( C ) c-kit + top (Z = 10) and ( D ) bottom (Z = 0). Confocal microscope, LEICA TCS SL, objective ×63, oil.

    Article Snippet: In the second series, the primary mouse anti-Isl1 (Abcam) and anti-Sca1 (Abcam) monoclonal antibodies were preliminary conjugated with Alexa 405 according to Zenon technology (Invitrogen) and then used for immunostaining at a 1:100 dilution.

    Techniques: Microscopy

    BMP signaling enhances Isl1-CPC cardiac differentiation. Freshly sorted EB day 5.5 GFP + Isl1-CPC were treated with control (A, D) or 25 ng/ml Bmp4 (B, E) for 6 days, followed by immunostaining with antibodies against cardiomyocyte-specific protein cTnT (red in A and B) or labelled with cTnT and Alexa fluor 647 for FACS analysis (D, E). Nuclei: blue (Hoechst). (C) Quantification of A and B (over 2000 cells counted from 3 independent experiments). (F) Quantification of D and E (20000 cells FACS analyzed for each of 3 independent experiments). Scale bar: 100 µm; *statistical significance p

    Journal: PLoS ONE

    Article Title: Functional Cardiomyocytes Derived from Isl1 Cardiac Progenitors via Bmp4 Stimulation

    doi: 10.1371/journal.pone.0110752

    Figure Lengend Snippet: BMP signaling enhances Isl1-CPC cardiac differentiation. Freshly sorted EB day 5.5 GFP + Isl1-CPC were treated with control (A, D) or 25 ng/ml Bmp4 (B, E) for 6 days, followed by immunostaining with antibodies against cardiomyocyte-specific protein cTnT (red in A and B) or labelled with cTnT and Alexa fluor 647 for FACS analysis (D, E). Nuclei: blue (Hoechst). (C) Quantification of A and B (over 2000 cells counted from 3 independent experiments). (F) Quantification of D and E (20000 cells FACS analyzed for each of 3 independent experiments). Scale bar: 100 µm; *statistical significance p

    Article Snippet: Immunofluorescence stainings were performed with mouse anti-Isl1 antibody (Hybridoma Bank) and rabbit anti-alpha smooth muscle actin (SMA from Abcam), cardiac troponin T (cTnT from Abcam) or CD31 (Abcam).

    Techniques: Immunostaining, FACS

    Isl1-CPC derived cultures have cardiomyocyte-like electrophysiological properties. Action potentials of differentiated Isl1-CPCs from control (A) or Bmp4-treated cultures (B) were recorded after 6 days in culture under whole-cell current clamp mode. (A, B) Representative action potentials demonstrate nodal-like, atrial-like and ventricular-like cardiomyocytes in the absence and presence of Bmp4. (C) Action potential amplitude (APA), action potential duration at the 90% of repolarization (APD 90 ) and maximum diastolic potential (MDP) for ventricular-like cardiomyocytes were quantified and depicted as bar graph for both control and Bmp4-treated cultures. A total of 29 cardiomyocytes derived from Isl1-CPCs was analyzed from 4 independent experiments. There were no significant differences for APA, MDP and APD 90 between control and Bmp4 treated samples. The details for AP measurement setting are under Materials and Methods .

    Journal: PLoS ONE

    Article Title: Functional Cardiomyocytes Derived from Isl1 Cardiac Progenitors via Bmp4 Stimulation

    doi: 10.1371/journal.pone.0110752

    Figure Lengend Snippet: Isl1-CPC derived cultures have cardiomyocyte-like electrophysiological properties. Action potentials of differentiated Isl1-CPCs from control (A) or Bmp4-treated cultures (B) were recorded after 6 days in culture under whole-cell current clamp mode. (A, B) Representative action potentials demonstrate nodal-like, atrial-like and ventricular-like cardiomyocytes in the absence and presence of Bmp4. (C) Action potential amplitude (APA), action potential duration at the 90% of repolarization (APD 90 ) and maximum diastolic potential (MDP) for ventricular-like cardiomyocytes were quantified and depicted as bar graph for both control and Bmp4-treated cultures. A total of 29 cardiomyocytes derived from Isl1-CPCs was analyzed from 4 independent experiments. There were no significant differences for APA, MDP and APD 90 between control and Bmp4 treated samples. The details for AP measurement setting are under Materials and Methods .

    Article Snippet: Immunofluorescence stainings were performed with mouse anti-Isl1 antibody (Hybridoma Bank) and rabbit anti-alpha smooth muscle actin (SMA from Abcam), cardiac troponin T (cTnT from Abcam) or CD31 (Abcam).

    Techniques: Derivative Assay

    Bmp4 signaling enhances transcription factor Tbx5 and Tbx20 mRNA expression. FACS-sorted GFP+ Isl1-CPCs were cultured in the presence of Bmp4 or vehicle control. mRNA was collected following 2, 4 and 6 days of Bmp4 treatment. qRT-PCR analysis of duplicates from three independent experiments for tbx5 (A) and tbx20 (B) expression is normalized by housekeeping gene, Gapdh. *p

    Journal: PLoS ONE

    Article Title: Functional Cardiomyocytes Derived from Isl1 Cardiac Progenitors via Bmp4 Stimulation

    doi: 10.1371/journal.pone.0110752

    Figure Lengend Snippet: Bmp4 signaling enhances transcription factor Tbx5 and Tbx20 mRNA expression. FACS-sorted GFP+ Isl1-CPCs were cultured in the presence of Bmp4 or vehicle control. mRNA was collected following 2, 4 and 6 days of Bmp4 treatment. qRT-PCR analysis of duplicates from three independent experiments for tbx5 (A) and tbx20 (B) expression is normalized by housekeeping gene, Gapdh. *p

    Article Snippet: Immunofluorescence stainings were performed with mouse anti-Isl1 antibody (Hybridoma Bank) and rabbit anti-alpha smooth muscle actin (SMA from Abcam), cardiac troponin T (cTnT from Abcam) or CD31 (Abcam).

    Techniques: Expressing, FACS, Cell Culture, Quantitative RT-PCR

    Calcium transients of cardiomyocytes derived from Isl1-CPC after cell loading with Ca 2+ indicator fura-2. (A) Representative recordings for increase in [Ca 2+ ]i transient amplitude and frequency was measured following loading of control and Bmp4-treated cardiomyocyte cultures with Ca 2+ indicator fura-2. Cytosolic Ca 2+ of spontaneous beating cardiomyocyte was measured by ratio of fluorescence intensity at 340 nm and 380 nm (F340/F380). The measurements were recorded 6 days of incubation after Isl1-CPC isolation. (B) The quantitative representation of beating frequency (BF) of calcium transients per minute is shown by bar graphs (n = 3). Experiments were repeated 3 times and 10–15 cells were measured in each experiment. (C) Change of [Ca 2+ ] was calculated from Ca 2+ transient recordings for cardiomyocyte cultures differentiated in the absence or presence of Bmp4. *p

    Journal: PLoS ONE

    Article Title: Functional Cardiomyocytes Derived from Isl1 Cardiac Progenitors via Bmp4 Stimulation

    doi: 10.1371/journal.pone.0110752

    Figure Lengend Snippet: Calcium transients of cardiomyocytes derived from Isl1-CPC after cell loading with Ca 2+ indicator fura-2. (A) Representative recordings for increase in [Ca 2+ ]i transient amplitude and frequency was measured following loading of control and Bmp4-treated cardiomyocyte cultures with Ca 2+ indicator fura-2. Cytosolic Ca 2+ of spontaneous beating cardiomyocyte was measured by ratio of fluorescence intensity at 340 nm and 380 nm (F340/F380). The measurements were recorded 6 days of incubation after Isl1-CPC isolation. (B) The quantitative representation of beating frequency (BF) of calcium transients per minute is shown by bar graphs (n = 3). Experiments were repeated 3 times and 10–15 cells were measured in each experiment. (C) Change of [Ca 2+ ] was calculated from Ca 2+ transient recordings for cardiomyocyte cultures differentiated in the absence or presence of Bmp4. *p

    Article Snippet: Immunofluorescence stainings were performed with mouse anti-Isl1 antibody (Hybridoma Bank) and rabbit anti-alpha smooth muscle actin (SMA from Abcam), cardiac troponin T (cTnT from Abcam) or CD31 (Abcam).

    Techniques: Derivative Assay, Fluorescence, Incubation, Isolation

    ISL1 directly regulates a number of genes required for normal pacemaker function in mice and human.

    Journal: The Journal of Clinical Investigation

    Article Title: Transcription factor ISL1 is essential for pacemaker development and function

    doi: 10.1172/JCI68257

    Figure Lengend Snippet: ISL1 directly regulates a number of genes required for normal pacemaker function in mice and human.

    Article Snippet: The following primary antibodies were used: mouse monoclonal anti-ISL1/2 (39.4D5, Developmental Studies Hybridoma Bank [DSHB]), rabbit anti-ISL1 (ab20670, Abcam), rat anti-HCN4 (ab32675, Abcam), rabbit anti-Cx40 (sc-28658, Santa Cruz), goat anti-TBX3 (sc-17871, Santa Cruz Biotechnology Inc.), and rat anti-BrdU (ab6326, Abcam).

    Techniques: Mouse Assay

    Bradycardia and reduced TBX3 and HCN4 expression following ablation of Isl1 during later SAN morphogenesis. ( A ) Ablation of Isl1 at E11.5 led to significantly slower heart rate at E12.5 and E14.5 ( n = 15 per group, P

    Journal: The Journal of Clinical Investigation

    Article Title: Transcription factor ISL1 is essential for pacemaker development and function

    doi: 10.1172/JCI68257

    Figure Lengend Snippet: Bradycardia and reduced TBX3 and HCN4 expression following ablation of Isl1 during later SAN morphogenesis. ( A ) Ablation of Isl1 at E11.5 led to significantly slower heart rate at E12.5 and E14.5 ( n = 15 per group, P

    Article Snippet: The following primary antibodies were used: mouse monoclonal anti-ISL1/2 (39.4D5, Developmental Studies Hybridoma Bank [DSHB]), rabbit anti-ISL1 (ab20670, Abcam), rat anti-HCN4 (ab32675, Abcam), rabbit anti-Cx40 (sc-28658, Santa Cruz), goat anti-TBX3 (sc-17871, Santa Cruz Biotechnology Inc.), and rat anti-BrdU (ab6326, Abcam).

    Techniques: Expressing

    Bradycardia and loss of SAN cells in Isl1 compound mutants. ISL1-nLacZ was expressed in SV myocardium, including the SAN region (red arrow), and mesocardium at E9.5 ( A and C ) and E11.5 ( G and I ). Expression of ISL1 and HCN4 in the SV region of Isl1 compound mutant embryos was significantly reduced ( E and F ). Expression of ISL1 and the number of ISL1-expressing cells in the SV, SAN (red arrow), and DM was markedly reduced in Isl1 compound mutant embryos at E9.5 ( B and D ) and E11.5 ( H and J ). BrdU staining revealed significantly reduced proliferation of SV myocardium in Isl1 compound mutants at E9.5 ( K – M ). TUNEL labeling showed significantly increased cell death in the SV of Isl1 compound mutant embryos at E10.5 ( N – P ) ( n = 4 per group. Scale bars as shown). Echocardiography revealed a significant reduction in the heart rate of Isl1 compound mutant embryos at E9.5 and E11.5 ( Q ). n = 15 per group; * P

    Journal: The Journal of Clinical Investigation

    Article Title: Transcription factor ISL1 is essential for pacemaker development and function

    doi: 10.1172/JCI68257

    Figure Lengend Snippet: Bradycardia and loss of SAN cells in Isl1 compound mutants. ISL1-nLacZ was expressed in SV myocardium, including the SAN region (red arrow), and mesocardium at E9.5 ( A and C ) and E11.5 ( G and I ). Expression of ISL1 and HCN4 in the SV region of Isl1 compound mutant embryos was significantly reduced ( E and F ). Expression of ISL1 and the number of ISL1-expressing cells in the SV, SAN (red arrow), and DM was markedly reduced in Isl1 compound mutant embryos at E9.5 ( B and D ) and E11.5 ( H and J ). BrdU staining revealed significantly reduced proliferation of SV myocardium in Isl1 compound mutants at E9.5 ( K – M ). TUNEL labeling showed significantly increased cell death in the SV of Isl1 compound mutant embryos at E10.5 ( N – P ) ( n = 4 per group. Scale bars as shown). Echocardiography revealed a significant reduction in the heart rate of Isl1 compound mutant embryos at E9.5 and E11.5 ( Q ). n = 15 per group; * P

    Article Snippet: The following primary antibodies were used: mouse monoclonal anti-ISL1/2 (39.4D5, Developmental Studies Hybridoma Bank [DSHB]), rabbit anti-ISL1 (ab20670, Abcam), rat anti-HCN4 (ab32675, Abcam), rabbit anti-Cx40 (sc-28658, Santa Cruz), goat anti-TBX3 (sc-17871, Santa Cruz Biotechnology Inc.), and rat anti-BrdU (ab6326, Abcam).

    Techniques: Expressing, Mutagenesis, BrdU Staining, TUNEL Assay, Labeling

    Reduced expression of Hcn4 , Tbx3 , and Shox2 in the SAN region of Isl1 compound mutant embryos. At E9.5, Hcn4 and Shox2 were expressed in the SV, and SAN region (red arrow; A , C , E , and G ). Tbx3 was expressed in the SV and surrounding mesenchyme (red arrow; I and K ). In Isl1 compound mutant embryos, expression of Hcn4, Shox2, and Tbx3 in the SV and SAN region was markedly reduced ( B , D , F , H , J , and L ). Cx40 and Nkx2-5 were expressed in working myocardium but not in the SAN region ( M , O , Q , and S In Isl1 compound mutant embryos, expression of Cx40 and Nkx2-5 was markedly reduced in atrial myocardium, but no expansion or ectopic expression of Cx40 or Nkx2-5 was observed in the SAN region ( N , P , R , and T ). n = 4 per group, Scale bars as shown.

    Journal: The Journal of Clinical Investigation

    Article Title: Transcription factor ISL1 is essential for pacemaker development and function

    doi: 10.1172/JCI68257

    Figure Lengend Snippet: Reduced expression of Hcn4 , Tbx3 , and Shox2 in the SAN region of Isl1 compound mutant embryos. At E9.5, Hcn4 and Shox2 were expressed in the SV, and SAN region (red arrow; A , C , E , and G ). Tbx3 was expressed in the SV and surrounding mesenchyme (red arrow; I and K ). In Isl1 compound mutant embryos, expression of Hcn4, Shox2, and Tbx3 in the SV and SAN region was markedly reduced ( B , D , F , H , J , and L ). Cx40 and Nkx2-5 were expressed in working myocardium but not in the SAN region ( M , O , Q , and S In Isl1 compound mutant embryos, expression of Cx40 and Nkx2-5 was markedly reduced in atrial myocardium, but no expansion or ectopic expression of Cx40 or Nkx2-5 was observed in the SAN region ( N , P , R , and T ). n = 4 per group, Scale bars as shown.

    Article Snippet: The following primary antibodies were used: mouse monoclonal anti-ISL1/2 (39.4D5, Developmental Studies Hybridoma Bank [DSHB]), rabbit anti-ISL1 (ab20670, Abcam), rat anti-HCN4 (ab32675, Abcam), rabbit anti-Cx40 (sc-28658, Santa Cruz), goat anti-TBX3 (sc-17871, Santa Cruz Biotechnology Inc.), and rat anti-BrdU (ab6326, Abcam).

    Techniques: Expressing, Mutagenesis

    Bradycardia and loss of SAN cells following ablation of Isl1 in SAN during early developmental stages using Hcn4-CreERT2 . Isl1 mutant ( Hcn4-CreERT2 Isl1 fl/fl ) and control ( Hcn4-CreERT2 Isl1 fl/+ or +/+ ) embryos were given tamoxifen at E9.5. Embryos were analyzed 36 and 48 hours after induction. ( A ) Echocardiography revealed that the heart rate of Isl1 mutants was significantly reduced at E11 and was further reduced at E11.5 ( n = 20 per group). ( B – D ) Whole-mount X-gal staining and quantitative analysis revealed a significantly reduced number of X-gal + and Tomato + cells in the SAN (red arrow) of Isl1 mutants relative to control littermates at E11.5 ( n = 4. Scale bars as shown). ( D – H ) Immunostaining demonstrated significantly reduced expression of HCN4 and TBX3 in the SAN of Isl1 mutants compared with controls marked by Tomato + at E11.5. However, a slight but not significant reduction in the number of Hcn4 lineage–labeled cells in Isl1 mutant SAN region was observed when analyzed at E11 ( D , I , and J ). ( K – M ) TUNEL revealed increased cell death in Isl1 mutant SAN marked by Tomato + . ( N – P ) BrdU revealed decreased proliferation in Isl1 mutant SAN marked by Tomato. n = 4; * P

    Journal: The Journal of Clinical Investigation

    Article Title: Transcription factor ISL1 is essential for pacemaker development and function

    doi: 10.1172/JCI68257

    Figure Lengend Snippet: Bradycardia and loss of SAN cells following ablation of Isl1 in SAN during early developmental stages using Hcn4-CreERT2 . Isl1 mutant ( Hcn4-CreERT2 Isl1 fl/fl ) and control ( Hcn4-CreERT2 Isl1 fl/+ or +/+ ) embryos were given tamoxifen at E9.5. Embryos were analyzed 36 and 48 hours after induction. ( A ) Echocardiography revealed that the heart rate of Isl1 mutants was significantly reduced at E11 and was further reduced at E11.5 ( n = 20 per group). ( B – D ) Whole-mount X-gal staining and quantitative analysis revealed a significantly reduced number of X-gal + and Tomato + cells in the SAN (red arrow) of Isl1 mutants relative to control littermates at E11.5 ( n = 4. Scale bars as shown). ( D – H ) Immunostaining demonstrated significantly reduced expression of HCN4 and TBX3 in the SAN of Isl1 mutants compared with controls marked by Tomato + at E11.5. However, a slight but not significant reduction in the number of Hcn4 lineage–labeled cells in Isl1 mutant SAN region was observed when analyzed at E11 ( D , I , and J ). ( K – M ) TUNEL revealed increased cell death in Isl1 mutant SAN marked by Tomato + . ( N – P ) BrdU revealed decreased proliferation in Isl1 mutant SAN marked by Tomato. n = 4; * P

    Article Snippet: The following primary antibodies were used: mouse monoclonal anti-ISL1/2 (39.4D5, Developmental Studies Hybridoma Bank [DSHB]), rabbit anti-ISL1 (ab20670, Abcam), rat anti-HCN4 (ab32675, Abcam), rabbit anti-Cx40 (sc-28658, Santa Cruz), goat anti-TBX3 (sc-17871, Santa Cruz Biotechnology Inc.), and rat anti-BrdU (ab6326, Abcam).

    Techniques: Mutagenesis, Staining, Immunostaining, Expressing, Labeling, TUNEL Assay

    RNA-seq analyses reveal dysregulation of a number of genes important for SAN function in Hcn4-CreERT2 Isl1 fl/fl mutants. ( A ) Scatterplot illustrating relative gene expression of polyA-selected RNA transcripts from RNA-seq comparison of control and Hcn4-CreERT2 Isl1 fl/fl mutant SAN cells. Genes upregulated or downregulated 1.5-fold in Isl1 mutant SAN cells are shown in red and green, respectively. Values are presented as log2 of tag counts normalized to 10 7 uniquely mapped tags. ( B ) RNA-seq comparison of control and Hcn4-CreERT2 Isl1 fl/fl mutant SAN transcriptomes revealed a total of 12,441 genes expressed (RPKM ≥ 1) in SAN cells, of which 1,035 upregulated and 3,690 downregulated in Isl1 mutant SAN cells ( |fold-change mutant vs. ctrl| ≥ 1.5). ( C ) GO functional clustering of genes down- and upregulated in Isl1 mutant, highlighting cellular processes most significantly affected in mutant SAN (top 10 not redundant categories are shown). ( D ) qPCR validation analysis. mRNA expression of ion channels and associated genes, and genes involved in transcription regulation, cell cycle, and signaling pathways are shown. ( E ) qRT-PCR validation analysis. mRNA expression of atrial myocardial specific genes. Results are shown as fold-change Isl1 mutant vs. ctrl. n = 4 per group, P

    Journal: The Journal of Clinical Investigation

    Article Title: Transcription factor ISL1 is essential for pacemaker development and function

    doi: 10.1172/JCI68257

    Figure Lengend Snippet: RNA-seq analyses reveal dysregulation of a number of genes important for SAN function in Hcn4-CreERT2 Isl1 fl/fl mutants. ( A ) Scatterplot illustrating relative gene expression of polyA-selected RNA transcripts from RNA-seq comparison of control and Hcn4-CreERT2 Isl1 fl/fl mutant SAN cells. Genes upregulated or downregulated 1.5-fold in Isl1 mutant SAN cells are shown in red and green, respectively. Values are presented as log2 of tag counts normalized to 10 7 uniquely mapped tags. ( B ) RNA-seq comparison of control and Hcn4-CreERT2 Isl1 fl/fl mutant SAN transcriptomes revealed a total of 12,441 genes expressed (RPKM ≥ 1) in SAN cells, of which 1,035 upregulated and 3,690 downregulated in Isl1 mutant SAN cells ( |fold-change mutant vs. ctrl| ≥ 1.5). ( C ) GO functional clustering of genes down- and upregulated in Isl1 mutant, highlighting cellular processes most significantly affected in mutant SAN (top 10 not redundant categories are shown). ( D ) qPCR validation analysis. mRNA expression of ion channels and associated genes, and genes involved in transcription regulation, cell cycle, and signaling pathways are shown. ( E ) qRT-PCR validation analysis. mRNA expression of atrial myocardial specific genes. Results are shown as fold-change Isl1 mutant vs. ctrl. n = 4 per group, P

    Article Snippet: The following primary antibodies were used: mouse monoclonal anti-ISL1/2 (39.4D5, Developmental Studies Hybridoma Bank [DSHB]), rabbit anti-ISL1 (ab20670, Abcam), rat anti-HCN4 (ab32675, Abcam), rabbit anti-Cx40 (sc-28658, Santa Cruz), goat anti-TBX3 (sc-17871, Santa Cruz Biotechnology Inc.), and rat anti-BrdU (ab6326, Abcam).

    Techniques: RNA Sequencing Assay, Expressing, Mutagenesis, Functional Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    Expression of ISL1 in pacemaker cells of the SAN during development and after birth. ISL1 was coexpressed with HCN4 in myocardium of the SV at E9.5 ( A ), and in the majority of SAN cells from E10.5–P7 ( B – G ). ISL1 expression did not overlap with Cx40, which is expressed in atrial myocardium ( E and G ). The boxed area in H delineates regions depicted in F and G . The fraction of HCN4 cells that expressed Isl1 remained constant at early stages from E11.5–E14.5, but decreased at E18 ( I ). After birth, the fraction of HCN4 cells that expressed Isl1 decreased significantly ( I ). n = 4, P

    Journal: The Journal of Clinical Investigation

    Article Title: Transcription factor ISL1 is essential for pacemaker development and function

    doi: 10.1172/JCI68257

    Figure Lengend Snippet: Expression of ISL1 in pacemaker cells of the SAN during development and after birth. ISL1 was coexpressed with HCN4 in myocardium of the SV at E9.5 ( A ), and in the majority of SAN cells from E10.5–P7 ( B – G ). ISL1 expression did not overlap with Cx40, which is expressed in atrial myocardium ( E and G ). The boxed area in H delineates regions depicted in F and G . The fraction of HCN4 cells that expressed Isl1 remained constant at early stages from E11.5–E14.5, but decreased at E18 ( I ). After birth, the fraction of HCN4 cells that expressed Isl1 decreased significantly ( I ). n = 4, P

    Article Snippet: The following primary antibodies were used: mouse monoclonal anti-ISL1/2 (39.4D5, Developmental Studies Hybridoma Bank [DSHB]), rabbit anti-ISL1 (ab20670, Abcam), rat anti-HCN4 (ab32675, Abcam), rabbit anti-Cx40 (sc-28658, Santa Cruz), goat anti-TBX3 (sc-17871, Santa Cruz Biotechnology Inc.), and rat anti-BrdU (ab6326, Abcam).

    Techniques: Expressing

    Cervical development. (A–D) are sagittal sections of the female reproductive tract of a 16 week fetus (A=H E stain, B=ISL1 immunostain, C–D=keratin 19 immunostain). Boundaries between the vagina, cervix and uterus are nebulous up to 18 weeks, when vaginal fornices become apparent (F). (B) ISL1 immunostaining is strong in vaginal stroma, absent in uterine stroma, with a sharp fall off in staining intensity at the mid-point of the uterovaginal canal (red arrow in B). Keratin 19 immunostaining (C–D) may also be indicative of vaginal-exocervical-endocervical boundaries. Cervical glands are prominent at 18 weeks of gestation (E).

    Journal: Differentiation; research in biological diversity

    Article Title: New insights insights into human female reproductive tract development

    doi: 10.1016/j.diff.2017.08.002

    Figure Lengend Snippet: Cervical development. (A–D) are sagittal sections of the female reproductive tract of a 16 week fetus (A=H E stain, B=ISL1 immunostain, C–D=keratin 19 immunostain). Boundaries between the vagina, cervix and uterus are nebulous up to 18 weeks, when vaginal fornices become apparent (F). (B) ISL1 immunostaining is strong in vaginal stroma, absent in uterine stroma, with a sharp fall off in staining intensity at the mid-point of the uterovaginal canal (red arrow in B). Keratin 19 immunostaining (C–D) may also be indicative of vaginal-exocervical-endocervical boundaries. Cervical glands are prominent at 18 weeks of gestation (E).

    Article Snippet: Selected paraffin sections were immunostained as described previously ( ) with the following antibodies: PAX2 (Abcam, Cambridge, MA, Catalogue # ab150391, 1/50), FOXA1 (Atlas Antibodies, Bromma, Sweden, Catalogue # HPA050505, 1/500), Isl1 (Abcam, Ab20670, 1/200), Keratin 19, E.B.

    Techniques: Staining, Immunostaining