Journal: Journal of Nuclear Cardiology

Article Title: Correction of collimator-dependent differences in the heart-to-mediastinum ratio in 123 I-metaiodobenzylguanidine cardiac sympathetic imaging: Determination of conversion equations using point-source imaging

doi: 10.1007/s12350-016-0546-8

Figure Lengend Snippet: Specifications of the collimators

Article Snippet: Combined correction is recommended for the Siemens LEHR, Siemens LEAP, Siemens SLEHR, and Philips CHR collimators if appropriate subwindow data are available, and empiric correction suffices for the Siemens LME collimator.

Techniques:

Journal: Journal of Nuclear Cardiology

Article Title: Correction of collimator-dependent differences in the heart-to-mediastinum ratio in 123 I-metaiodobenzylguanidine cardiac sympathetic imaging: Determination of conversion equations using point-source imaging

doi: 10.1007/s12350-016-0546-8

Figure Lengend Snippet: Patient-based correction of the H/M ratios obtained with non-ME ( A , LEAP; B , SLEHR; C , LME; D , CHR) collimators. The values before correction ( green square ), after empiric correction ( blue circle ), and after combined correction ( red triangle ) were plotted against the uncorrected values obtained with the ME collimator. The broken line indicates the line of identity. Results of linear regression are shown

Article Snippet: Combined correction is recommended for the Siemens LEHR, Siemens LEAP, Siemens SLEHR, and Philips CHR collimators if appropriate subwindow data are available, and empiric correction suffices for the Siemens LME collimator.

Techniques:

Journal: Journal of Nuclear Cardiology

Article Title: Correction of collimator-dependent differences in the heart-to-mediastinum ratio in 123 I-metaiodobenzylguanidine cardiac sympathetic imaging: Determination of conversion equations using point-source imaging

doi: 10.1007/s12350-016-0546-8

Figure Lengend Snippet: Errors before and after patient-based correction ( A , LEAP; B , SLEHR; C , LME; D , CHR). Errors before correction ( green square ), after empiric correction ( blue circle ), and after combined correction ( red triangle ) were plotted against the uncorrected H/M ratios obtained with the ME collimator. Results of linear regression are shown

Article Snippet: Combined correction is recommended for the Siemens LEHR, Siemens LEAP, Siemens SLEHR, and Philips CHR collimators if appropriate subwindow data are available, and empiric correction suffices for the Siemens LME collimator.

Techniques:

Journal: Journal of Nuclear Cardiology

Article Title: Correction of collimator-dependent differences in the heart-to-mediastinum ratio in 123 I-metaiodobenzylguanidine cardiac sympathetic imaging: Determination of conversion equations using point-source imaging

doi: 10.1007/s12350-016-0546-8

Figure Lengend Snippet: Point-source-based correction of the H/M ratios obtained with non-ME ( A , LEAP; B , SLEHR; C , LME; D , CHR) collimators. The values after empiric correction ( blue circle ) and after combined correction ( red triangle ) were plotted against the uncorrected values obtained with the ME collimator. The broken line indicates the line of identity. Results of linear regression are shown

Article Snippet: Combined correction is recommended for the Siemens LEHR, Siemens LEAP, Siemens SLEHR, and Philips CHR collimators if appropriate subwindow data are available, and empiric correction suffices for the Siemens LME collimator.

Techniques:

Journal: Journal of Nuclear Cardiology

Article Title: Correction of collimator-dependent differences in the heart-to-mediastinum ratio in 123 I-metaiodobenzylguanidine cardiac sympathetic imaging: Determination of conversion equations using point-source imaging

doi: 10.1007/s12350-016-0546-8

Figure Lengend Snippet: Errors after point-source-based correction ( A , LEAP; B , SLEHR; C , LME; D , CHR) collimators. Errors after empiric correction ( blue circle ) and combined correction ( red triangle ) were plotted against the uncorrected H/M ratios obtained with the ME collimator. Results of linear regression are shown

Article Snippet: Combined correction is recommended for the Siemens LEHR, Siemens LEAP, Siemens SLEHR, and Philips CHR collimators if appropriate subwindow data are available, and empiric correction suffices for the Siemens LME collimator.

Techniques:

Journal: Nature Communications

Article Title: Bidirectional Wnt signaling between endoderm and mesoderm confers tracheal identity in mouse and human cells

doi: 10.1038/s41467-020-17969-w

Figure Lengend Snippet: a Schematic model of tracheoesophageal segregation. b Transverse sections of Nkx2.1 null mouse embryos and littermate controls. Sections were stained for Sox2 ( green ), Tbx4 ( magenta ), and DAPI ( blue ). Arrowheads indicate Tbx4 + tracheal mesoderm. Asterisks indicate nonspecific background signal of blood cells in dorsal aorta. n = 3/3 embryos per genotype. c Transverse sections of Shh Cre , Ctnnb1 flox/flox mouse embryos and littermate controls. Sections were stained by Sox2 ( green ), Tbx4 ( magenta ), and DAPI ( blue ). Arrowheads indicate Tbx4 + tracheal mesoderm. n = 3/3 embryos per genotype. d Transverse sections of Shh Cre , Ctnnb1 flox/flox mouse embryos-, and littermate controls. Sections were stained by Nkx2.1 ( magenta ) and DAPI ( blue ). n = 3/3 embryos per genotype. n neural tube, a aorta, Es Esophagus, Tr Trachea, Tr–E Trachea–Esophageal tube. Scale bar, 40 μm.

Article Snippet: The fraction of mouse Tbx4 lung mesenchyme specific enhancer (LME) (mm10, chr11:85,893,703-85,894,206, GenScript, ID U3154EL200-3) or Tbx4 -LME containing putative Tcf/Lef sites mutated (GenScript, ID U3154EL200-6) were synthesized and cloned into pGL4.23 (luc2/minP) vector (promega). mESC-derived LPM cells were transfected at day 5 in 150 μl of Opti-MEM (Thermo Fisher Scientific, 31985088) with 2 μl of Lipofectamine Stem (Thermo Fisher Scientific, STEM00003) and 1 μg of pGL4.23 (luc2/minP) containing a fraction of mouse Tbx4 -LME or Tbx4 -LME containing mutated Tcf/Lef sites.

Techniques: Staining

Journal: Nature Communications

Article Title: Bidirectional Wnt signaling between endoderm and mesoderm confers tracheal identity in mouse and human cells

doi: 10.1038/s41467-020-17969-w

Figure Lengend Snippet: a Transverse sections of LEF1 EGFP reporter mouse embryos at E9.5 to E11.5. Sections were stained for EGFP ( green ), Nkx2.1 ( magenta ), and DAPI ( blue ). Arrowheads indicate GFP + cells. n = 3/3 embryos. b Transverse sections of LEF1 EGFP reporter mouse embryos at E9.5 to E11.5. Sections were stained for EGFP ( green ), Tbx4 ( magenta ), and DAPI ( blue ). Arrowheads indicate GFP + cells. n = 3/3 embryos per genotype. c Transversal section of mouse embryo at E10.5. Section were stained for Axin2 ( green ), Nkx2.1 ( magenta ) and DAPI ( blue ) by RNAscope experiment. Arrowheads indicate Axin2 + cells. n = 2/2 embryos. d Transverse sections of Dermo1 Cre , Ctnnb1 flox/flox mouse embryos and littermate controls at E10.5. Upper panels show sections stained for Sox2 ( green ), Tbx4 ( magenta ), and DAPI ( blue ). Lower panels show sections stained for Nkx2.1 ( magenta ) and DAPI ( blue ). Arrowhead indicates Tbx4 + cells. Arrows indicate Nkx2.1 + cells. n = 3/3 embryos per genotype. e Three-dimensional imaging of whole trachea and esophagus tissue at E16.5. Cartilage morphology and smooth muscle architecture in the tracheas of Dermo1 Cre , Ctnnb1 flox/flox mouse embryos and control littermates. Whole trachea and esophagus were stained for Sox9 ( green ) and SMA ( magenta ). n = 3/3 embryos per genotype. f Model of tracheal architecture in Dermo1 Cre , Ctnnb1 flox/flox mouse embryos and control littermates based on e . g Integrative Genomics Viewer (IGV) snapshot of mm10 (chr11:85,884,521-85,904,766) showing mouse tbx4 lung mesenchyme specific element (LME) and compiled ENCODE data of ATAC-seq E14.5 lung (ENCSR335VJW), H3K27Ac E14.5 lung (ENCSR452WYC), H3K4me1 E15.5 lung (ENCFF283EBS), EP300 postnatal day (PND) 0 lung and vertebrate conservation (Phastcons). Numbers indicate fold enrichment over input (ChIP-seq). CisBP and Jaspar predicted Tcf/Lef-binding sites (highlighted in green, region: mm10, chr11:85,893,703-85,894,206) are localized at the ATAC-seq and p300 peaks that are conserved among most vertebrates. Sequence in red shows the Tcf/Lef-binding sites mutated. Eso Esophagus, Lu Lung, Tr Trachea, Tr–E Tracheoesophageal tube. Scale bar: 40 μm ( a , b ), 50 μm ( c , d ), 300 μm ( e ).

Article Snippet: The fraction of mouse Tbx4 lung mesenchyme specific enhancer (LME) (mm10, chr11:85,893,703-85,894,206, GenScript, ID U3154EL200-3) or Tbx4 -LME containing putative Tcf/Lef sites mutated (GenScript, ID U3154EL200-6) were synthesized and cloned into pGL4.23 (luc2/minP) vector (promega). mESC-derived LPM cells were transfected at day 5 in 150 μl of Opti-MEM (Thermo Fisher Scientific, 31985088) with 2 μl of Lipofectamine Stem (Thermo Fisher Scientific, STEM00003) and 1 μg of pGL4.23 (luc2/minP) containing a fraction of mouse Tbx4 -LME or Tbx4 -LME containing mutated Tcf/Lef sites.

Techniques: Staining, RNAscope, Imaging, Control, ChIP-sequencing, Binding Assay, Sequencing

Journal: Nature Communications

Article Title: Bidirectional Wnt signaling between endoderm and mesoderm confers tracheal identity in mouse and human cells

doi: 10.1038/s41467-020-17969-w

Figure Lengend Snippet: a In situ hybridization for Wnt2 mRNA during tracheoesophageal segregation. Arrowheads indicate Wnt2 expression in the ventrolateral mesoderm at E9.5 and E10.5. n = 2/2 embryos. b Transverse sections of Shh Cre , Wls flox/flox mouse embryos and littermate controls at E10.5. Left panels show sections stained with Sox2 ( green ), Tbx4 ( magenta ), and DAPI ( blue ). Right panels show sections stained for Nkx2.1 ( magenta ) and DAPI ( blue ). n = 3/3 embryos per genotype. c In situ hybridization for Wnt2 mRNA in Shh Cre , Wls flox/flox mouse embryos and littermate controls at E9.5. Arrowheads indicate Wnt2 expression in the ventrolateral mesoderm. n = 2/2 embryos. d Refined model of tracheoesophageal segregation and tracheal mesodermal differentiation. e In situ hybridization for Wnt7b mRNA in mouse embryo at E10.5. Arrowhead indicates Wnt7b + cells. n = 2/2 embryos. Eso Esophagus, Lu Lung, Tr Trachea. Scale bar; 40 μm ( a – c ), 50 μm ( e ).

Article Snippet: The fraction of mouse Tbx4 lung mesenchyme specific enhancer (LME) (mm10, chr11:85,893,703-85,894,206, GenScript, ID U3154EL200-3) or Tbx4 -LME containing putative Tcf/Lef sites mutated (GenScript, ID U3154EL200-6) were synthesized and cloned into pGL4.23 (luc2/minP) vector (promega). mESC-derived LPM cells were transfected at day 5 in 150 μl of Opti-MEM (Thermo Fisher Scientific, 31985088) with 2 μl of Lipofectamine Stem (Thermo Fisher Scientific, STEM00003) and 1 μg of pGL4.23 (luc2/minP) containing a fraction of mouse Tbx4 -LME or Tbx4 -LME containing mutated Tcf/Lef sites.

Techniques: In Situ Hybridization, Expressing, Staining

Journal: Nature Communications

Article Title: Bidirectional Wnt signaling between endoderm and mesoderm confers tracheal identity in mouse and human cells

doi: 10.1038/s41467-020-17969-w

Figure Lengend Snippet: a Experimental design to generate tracheal mesoderm from mESCs. b Differentiating cells from mESCs at day 5. Cells were stained for Foxf1 ( magenta ) and Gata4 ( green ), respectively. % was calculated from randomly chosen 3 fields. Images are representative of two independent experiments. c qRT-PCR for LPM markers of mESC-derived LPM ( n = 3 independent wells). Experiments were repeated at least twice. d Differentiating cells from mESCs at day 6. Cells were stained for Tbx4 ( green ) and Foxf1 ( magenta ). % was calculated from randomly chosen three fields. Images are representative of at least two independent experiments. e qRT-PCR for respiratory mesoderm marker expression of mESC-derived trachea mesodermal cells. ( n = 3 independent wells). Experiments were repeated at least twice. f Diagram showing the constructs utilized in luciferase experiments containing Tbx4- − − LME Tbx4-LME wild-type containing five Tcf/Lef-binding sites and Mutant. mESC-derived LPMs were transfected with wild-type or mutant Tbx4 -LME during 4hrs following by respiratory induction in presence or absence of CHIR99021. g Luciferase assay examining the activation of Tbx4 -LME wt and mutant in response to 3 μM CHIR99021. P -values were provided by two-sided Tukey’s multiple comparison. *** p < 0.0001 ( n = 3 from independent wells from a single experiment). h Differentiating cells from mESCs at day 12. Cells were stained for Acta2 ( magenta ) and Sox9 ( green ). The asterisk indicates Sox9 + /SMA - chondrocyte aggregates. Images are representative of two independent experiments. i qRT-PCR for Sox9 and Acta2 expression of hESC-derived trachea mesodermal cells ( n = 3 independent wells). Experiments were repeated twice. j Differentiating cells from mESCs at day 12. Chondrocytes were stained with Alcian blue. The asterisk indicates one of the chondrocyte aggregates. Images are representative of two independent experiments. k Differentiating cells from mESCs at day 12. Cells were stained for Col1a1 ( magenta ) and Aggrecan ( green ). Image is representative of two independent experiments. l Differentiating cells from mESCs at day 12. Cells were stained for Tagln ( magenta ) and Col2a1 ( green ). Image is representative of two independent experiments. Each column shows the mean with S.D. Scale bar; 50 μm. Source data for b, c, d, e, g, i are provided in Source data file.

Article Snippet: The fraction of mouse Tbx4 lung mesenchyme specific enhancer (LME) (mm10, chr11:85,893,703-85,894,206, GenScript, ID U3154EL200-3) or Tbx4 -LME containing putative Tcf/Lef sites mutated (GenScript, ID U3154EL200-6) were synthesized and cloned into pGL4.23 (luc2/minP) vector (promega). mESC-derived LPM cells were transfected at day 5 in 150 μl of Opti-MEM (Thermo Fisher Scientific, 31985088) with 2 μl of Lipofectamine Stem (Thermo Fisher Scientific, STEM00003) and 1 μg of pGL4.23 (luc2/minP) containing a fraction of mouse Tbx4 -LME or Tbx4 -LME containing mutated Tcf/Lef sites.

Techniques: Staining, Quantitative RT-PCR, Derivative Assay, Marker, Expressing, Construct, Luciferase, Binding Assay, Mutagenesis, Transfection, Activation Assay, Comparison

Journal: Nature Communications

Article Title: Bidirectional Wnt signaling between endoderm and mesoderm confers tracheal identity in mouse and human cells

doi: 10.1038/s41467-020-17969-w

Figure Lengend Snippet: a Experimental design to generate tracheal mesoderm from hESCs. b Differentiating cells from hESCs at day 2. Cells were stained for FOXF1 ( magenta ) and GATA4 ( green ), respectively. % was calculated from randomly chosen 3 fields. Images are representative of two experiments. c qRT-PCR for LPM marker expression of hESC-derived LPM ( n = 3 independent wells). Experiments were repeated at least twice. d Differentiating cells from hESCs at day 5. Cells were stained for TBX4 ( green ) and FOXF1 ( magenta ). % was calculated from randomly chosen three fields. Images are representative of three wells in a single experiment. e qRT-PCR for respiratory mesoderm marker expression of hESC-derived trachea mesodermal cells ( n = 3 independent wells). Experiments were repeated at least twice. f Differentiating cells from hESCs at day 10. Cells were stained for TBX4 ( green ) and NKX6.1 ( magenta ). Images are representative of three wells in a single experiment. White arrows; TBX4 + /NKX6.1 + mesodermal cells. White arrowheads; TBX4 + /NKX6.1 − mesodermal cells. Grey arrows; TBX4 − /NKX6.1 + mesodermal cells. g The rate of differentiated cells at day 10. % was calculated from randomly chosen three fields. Images are representative of three wells in a single experiment. h Differentiating cells from hESCs with or without CHIR99021 at day 10. Cells were stained for ACTA2 ( magenta ) and SOX9 ( green ). Images are representative of at least two experiments. i qRT-PCR for SOX9 and ACTA2 expression of hESC-derived trachea mesodermal cells with different doses of CHIR99021 ( n = 3 independent well). Experiments were repeated twice. j Differentiating cells from hESCs at day 10. Chondrocytes were stained with Alcian blue. The asterisk indicates a chondrocyte aggregate. Images are representative of two experiments. k Differentiating cells from hESCs at day 10. Cells were stained for COL1A1 ( magenta ) and AGGRECAN ( green ). Images are representative of two experiments. l Differentiating cells from hESCs at day 10. Cells were stained for TAGLN ( magenta ) and COL2A1 ( green ). Images are representative of two experiments. Each column shows the mean with S.D. ( n = 3). Scale bar; 50 μm ( d, f, h, j, k, l ), 100 μm ( b ). Source data for b, c, d, e, g, i are provided in Source data file.

Article Snippet: The fraction of mouse Tbx4 lung mesenchyme specific enhancer (LME) (mm10, chr11:85,893,703-85,894,206, GenScript, ID U3154EL200-3) or Tbx4 -LME containing putative Tcf/Lef sites mutated (GenScript, ID U3154EL200-6) were synthesized and cloned into pGL4.23 (luc2/minP) vector (promega). mESC-derived LPM cells were transfected at day 5 in 150 μl of Opti-MEM (Thermo Fisher Scientific, 31985088) with 2 μl of Lipofectamine Stem (Thermo Fisher Scientific, STEM00003) and 1 μg of pGL4.23 (luc2/minP) containing a fraction of mouse Tbx4 -LME or Tbx4 -LME containing mutated Tcf/Lef sites.

Techniques: Staining, Quantitative RT-PCR, Marker, Expressing, Derivative Assay