cripto Search Results


cripto  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc cripto
Cripto, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse cripto antibody  (R&D Systems)
93
R&D Systems mouse cripto antibody
The production yield, activity, and purification <t>of</t> <t>recombinant</t> <t>Cripto</t> produced in the 3D microcarriers as compared to the 2D method. ( A ) SDS-PAGE analysis of the purification steps of recombinant Cripto: lane M is the protein molecular weight marker; lane 1 is the protein solution from ultrafiltration; lane 2 is the flowthrough after the first passage through the His-tag affinity Ni-NTA resin; lane 3 is the flowthrough after the second passage through the same resin; lane 4 is the first wash step; lane 5 is the second wash step; lane 6 is the first elution from the His-tag affinity Ni-NTA resin; and lane 7 is the second elution from the same resin. The band at approximately 27 kDa is the Cripto protein (indicated by the red arrow). ( B ) Quantitative amounts of Cripto protein produced in the 3D batch (Cripto (3D) ) with HEK293 cells encapsulated in PF microcarriers and incubated in bioreactors after three rounds of harvesting are compared to the maximum amount of Cripto produced in the 2D batch (Cripto (2D) ) with HEK293 cells adherent to cell culture plates and cultured to their density threshold limits. An initial cell seeding of 3.2 × 10 6 cells was used for both techniques. ( C ) The biological activity of recombinant Cripto was compared for Cripto produced in PF microcarriers versus the 2D method by measuring binding affinity to the AlK4 receptor. Four independent experiments were carried out for the 3D system and three independent experiments were performed for the 2D cultivation method. Results are shown as mean ± S.D. *** indicates p < 0.001.
Mouse Cripto Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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recombinant mouse cripto 1 rmcr 1 proteins  (R&D Systems)
91
R&D Systems recombinant mouse cripto 1 rmcr 1 proteins
The production yield, activity, and purification <t>of</t> <t>recombinant</t> <t>Cripto</t> produced in the 3D microcarriers as compared to the 2D method. ( A ) SDS-PAGE analysis of the purification steps of recombinant Cripto: lane M is the protein molecular weight marker; lane 1 is the protein solution from ultrafiltration; lane 2 is the flowthrough after the first passage through the His-tag affinity Ni-NTA resin; lane 3 is the flowthrough after the second passage through the same resin; lane 4 is the first wash step; lane 5 is the second wash step; lane 6 is the first elution from the His-tag affinity Ni-NTA resin; and lane 7 is the second elution from the same resin. The band at approximately 27 kDa is the Cripto protein (indicated by the red arrow). ( B ) Quantitative amounts of Cripto protein produced in the 3D batch (Cripto (3D) ) with HEK293 cells encapsulated in PF microcarriers and incubated in bioreactors after three rounds of harvesting are compared to the maximum amount of Cripto produced in the 2D batch (Cripto (2D) ) with HEK293 cells adherent to cell culture plates and cultured to their density threshold limits. An initial cell seeding of 3.2 × 10 6 cells was used for both techniques. ( C ) The biological activity of recombinant Cripto was compared for Cripto produced in PF microcarriers versus the 2D method by measuring binding affinity to the AlK4 receptor. Four independent experiments were carried out for the 3D system and three independent experiments were performed for the 2D cultivation method. Results are shown as mean ± S.D. *** indicates p < 0.001.
Recombinant Mouse Cripto 1 Rmcr 1 Proteins, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse anti hcr1 moab  (R&D Systems)
92
R&D Systems mouse anti hcr1 moab
The production yield, activity, and purification <t>of</t> <t>recombinant</t> <t>Cripto</t> produced in the 3D microcarriers as compared to the 2D method. ( A ) SDS-PAGE analysis of the purification steps of recombinant Cripto: lane M is the protein molecular weight marker; lane 1 is the protein solution from ultrafiltration; lane 2 is the flowthrough after the first passage through the His-tag affinity Ni-NTA resin; lane 3 is the flowthrough after the second passage through the same resin; lane 4 is the first wash step; lane 5 is the second wash step; lane 6 is the first elution from the His-tag affinity Ni-NTA resin; and lane 7 is the second elution from the same resin. The band at approximately 27 kDa is the Cripto protein (indicated by the red arrow). ( B ) Quantitative amounts of Cripto protein produced in the 3D batch (Cripto (3D) ) with HEK293 cells encapsulated in PF microcarriers and incubated in bioreactors after three rounds of harvesting are compared to the maximum amount of Cripto produced in the 2D batch (Cripto (2D) ) with HEK293 cells adherent to cell culture plates and cultured to their density threshold limits. An initial cell seeding of 3.2 × 10 6 cells was used for both techniques. ( C ) The biological activity of recombinant Cripto was compared for Cripto produced in PF microcarriers versus the 2D method by measuring binding affinity to the AlK4 receptor. Four independent experiments were carried out for the 3D system and three independent experiments were performed for the 2D cultivation method. Results are shown as mean ± S.D. *** indicates p < 0.001.
Mouse Anti Hcr1 Moab, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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recombinant soluble human cr1  (R&D Systems)
90
R&D Systems recombinant soluble human cr1
FIGURE 1. Characterization of the recom- binant <t>CR1</t> CCP 22–30_His fragment. (A) SDS-PAGE analysis of 4 mg CR1 CCP22-30 under reducing (R) and nonreducing (NR) conditions. The positions of the m.w. mark- ers are indicated. (B) CD spectroscopy of CR1 CCP22–30. A spectrum was recorded in the far-UV (200–260 nm) and collected six times. The mean values for each wavelength were calculated. The maximal ellipticity is indicated by an arrowhead. (C) Electron microscopy analysis of CR1 CCP22–30 (375000) after negative staining with 2% sodium silicotungstate.
Recombinant Soluble Human Cr1, supplied by R&D Systems, 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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cripto  (Rockland Immunochemicals)
92
Rockland Immunochemicals cripto
FIGURE 1. Characterization of the recom- binant <t>CR1</t> CCP 22–30_His fragment. (A) SDS-PAGE analysis of 4 mg CR1 CCP22-30 under reducing (R) and nonreducing (NR) conditions. The positions of the m.w. mark- ers are indicated. (B) CD spectroscopy of CR1 CCP22–30. A spectrum was recorded in the far-UV (200–260 nm) and collected six times. The mean values for each wavelength were calculated. The maximal ellipticity is indicated by an arrowhead. (C) Electron microscopy analysis of CR1 CCP22–30 (375000) after negative staining with 2% sodium silicotungstate.
Cripto, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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antibodies mabs  (R&D Systems)
91
R&D Systems antibodies mabs
FIGURE 1. Lipid raft localization and release of CR-1 protein. A, NTERA2/D1 cells were stained with anti-CR-1 <t>mAb</t> (green), and lipid raft marker GM-1 was labeled with CTxB (red) without permeabilization and analyzed by confocal microscopy (upper panel). Magnified images for the marked regions in the upper panel are shown in the lower left panel. Negative controls without pri- mary antibody are shown in the lower right panel. Nuclei were stained with DAPI (blue) in all merged images. Scale bar 10 m. B, biochemical isolation of DRMs with 1% Triton X-100 by sucrose-gradient centrifugation of NTERA2/D1 cells. The gradient fractions (lanes 4–12) were analyzed with Western blot analysis for CR-1. Transferrin receptor (TfR) was used as a control for non-raft membrane protein. The lipid raft marker GM-1 in the same sam- ples was detected with HRP-conjugated CTxB by dot-blot analysis. C, compar- ative study of cell-associated (CL) and released (CM) CR-1 with or without serum by semi-quantitative Western blot analysis (see Table 1 and supple- mental Fig. S1, A–C). A representative blot for NTERA2/D1 cells is shown. The indicated amount of recombinant human CR-1 protein was used as a standard.
Antibodies Mabs, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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recombinant human cripto  (R&D Systems)
90
R&D Systems recombinant human cripto
FIGURE 1. Lipid raft localization and release of CR-1 protein. A, NTERA2/D1 cells were stained with anti-CR-1 <t>mAb</t> (green), and lipid raft marker GM-1 was labeled with CTxB (red) without permeabilization and analyzed by confocal microscopy (upper panel). Magnified images for the marked regions in the upper panel are shown in the lower left panel. Negative controls without pri- mary antibody are shown in the lower right panel. Nuclei were stained with DAPI (blue) in all merged images. Scale bar 10 m. B, biochemical isolation of DRMs with 1% Triton X-100 by sucrose-gradient centrifugation of NTERA2/D1 cells. The gradient fractions (lanes 4–12) were analyzed with Western blot analysis for CR-1. Transferrin receptor (TfR) was used as a control for non-raft membrane protein. The lipid raft marker GM-1 in the same sam- ples was detected with HRP-conjugated CTxB by dot-blot analysis. C, compar- ative study of cell-associated (CL) and released (CM) CR-1 with or without serum by semi-quantitative Western blot analysis (see Table 1 and supple- mental Fig. S1, A–C). A representative blot for NTERA2/D1 cells is shown. The indicated amount of recombinant human CR-1 protein was used as a standard.
Recombinant Human Cripto, supplied by R&D Systems, 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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sheep anti mouse cripto ab  (R&D Systems)
93
R&D Systems sheep anti mouse cripto ab
FIGURE 1. Lipid raft localization and release of CR-1 protein. A, NTERA2/D1 cells were stained with anti-CR-1 <t>mAb</t> (green), and lipid raft marker GM-1 was labeled with CTxB (red) without permeabilization and analyzed by confocal microscopy (upper panel). Magnified images for the marked regions in the upper panel are shown in the lower left panel. Negative controls without pri- mary antibody are shown in the lower right panel. Nuclei were stained with DAPI (blue) in all merged images. Scale bar 10 m. B, biochemical isolation of DRMs with 1% Triton X-100 by sucrose-gradient centrifugation of NTERA2/D1 cells. The gradient fractions (lanes 4–12) were analyzed with Western blot analysis for CR-1. Transferrin receptor (TfR) was used as a control for non-raft membrane protein. The lipid raft marker GM-1 in the same sam- ples was detected with HRP-conjugated CTxB by dot-blot analysis. C, compar- ative study of cell-associated (CL) and released (CM) CR-1 with or without serum by semi-quantitative Western blot analysis (see Table 1 and supple- mental Fig. S1, A–C). A representative blot for NTERA2/D1 cells is shown. The indicated amount of recombinant human CR-1 protein was used as a standard.
Sheep Anti Mouse Cripto Ab, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cripto 1  (Novus Biologicals)
90
Novus Biologicals cripto 1
FIGURE 1. Lipid raft localization and release of CR-1 protein. A, NTERA2/D1 cells were stained with anti-CR-1 <t>mAb</t> (green), and lipid raft marker GM-1 was labeled with CTxB (red) without permeabilization and analyzed by confocal microscopy (upper panel). Magnified images for the marked regions in the upper panel are shown in the lower left panel. Negative controls without pri- mary antibody are shown in the lower right panel. Nuclei were stained with DAPI (blue) in all merged images. Scale bar 10 m. B, biochemical isolation of DRMs with 1% Triton X-100 by sucrose-gradient centrifugation of NTERA2/D1 cells. The gradient fractions (lanes 4–12) were analyzed with Western blot analysis for CR-1. Transferrin receptor (TfR) was used as a control for non-raft membrane protein. The lipid raft marker GM-1 in the same sam- ples was detected with HRP-conjugated CTxB by dot-blot analysis. C, compar- ative study of cell-associated (CL) and released (CM) CR-1 with or without serum by semi-quantitative Western blot analysis (see Table 1 and supple- mental Fig. S1, A–C). A representative blot for NTERA2/D1 cells is shown. The indicated amount of recombinant human CR-1 protein was used as a standard.
Cripto 1, supplied by Novus Biologicals, 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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goat anti human cripto 1  (R&D Systems)
90
R&D Systems goat anti human cripto 1
FIGURE 1. Lipid raft localization and release of CR-1 protein. A, NTERA2/D1 cells were stained with anti-CR-1 <t>mAb</t> (green), and lipid raft marker GM-1 was labeled with CTxB (red) without permeabilization and analyzed by confocal microscopy (upper panel). Magnified images for the marked regions in the upper panel are shown in the lower left panel. Negative controls without pri- mary antibody are shown in the lower right panel. Nuclei were stained with DAPI (blue) in all merged images. Scale bar 10 m. B, biochemical isolation of DRMs with 1% Triton X-100 by sucrose-gradient centrifugation of NTERA2/D1 cells. The gradient fractions (lanes 4–12) were analyzed with Western blot analysis for CR-1. Transferrin receptor (TfR) was used as a control for non-raft membrane protein. The lipid raft marker GM-1 in the same sam- ples was detected with HRP-conjugated CTxB by dot-blot analysis. C, compar- ative study of cell-associated (CL) and released (CM) CR-1 with or without serum by semi-quantitative Western blot analysis (see Table 1 and supple- mental Fig. S1, A–C). A representative blot for NTERA2/D1 cells is shown. The indicated amount of recombinant human CR-1 protein was used as a standard.
Goat Anti Human Cripto 1, supplied by R&D Systems, 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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resource source identifier antibodies mouse cripto allophycocyanin mab biotechne r d systems  (R&D Systems)
93
R&D Systems resource source identifier antibodies mouse cripto allophycocyanin mab biotechne r d systems
Figure 1. <t>CRIPTO</t> micro-heterogeneity in ASC population (A) Schematic representation of the experimental design. (B) Representative FACS plots showing the gating strategy. The percentage of CRIPTO-positive cells above the threshold is indicated (bottom panel).
Resource Source Identifier Antibodies Mouse Cripto Allophycocyanin Mab Biotechne R D Systems, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/resource source identifier antibodies mouse cripto allophycocyanin mab biotechne r d systems/product/R&D Systems
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Image Search Results


The production yield, activity, and purification of recombinant Cripto produced in the 3D microcarriers as compared to the 2D method. ( A ) SDS-PAGE analysis of the purification steps of recombinant Cripto: lane M is the protein molecular weight marker; lane 1 is the protein solution from ultrafiltration; lane 2 is the flowthrough after the first passage through the His-tag affinity Ni-NTA resin; lane 3 is the flowthrough after the second passage through the same resin; lane 4 is the first wash step; lane 5 is the second wash step; lane 6 is the first elution from the His-tag affinity Ni-NTA resin; and lane 7 is the second elution from the same resin. The band at approximately 27 kDa is the Cripto protein (indicated by the red arrow). ( B ) Quantitative amounts of Cripto protein produced in the 3D batch (Cripto (3D) ) with HEK293 cells encapsulated in PF microcarriers and incubated in bioreactors after three rounds of harvesting are compared to the maximum amount of Cripto produced in the 2D batch (Cripto (2D) ) with HEK293 cells adherent to cell culture plates and cultured to their density threshold limits. An initial cell seeding of 3.2 × 10 6 cells was used for both techniques. ( C ) The biological activity of recombinant Cripto was compared for Cripto produced in PF microcarriers versus the 2D method by measuring binding affinity to the AlK4 receptor. Four independent experiments were carried out for the 3D system and three independent experiments were performed for the 2D cultivation method. Results are shown as mean ± S.D. *** indicates p < 0.001.

Journal: Gels

Article Title: Biomanufacturing Recombinantly Expressed Cripto-1 Protein in Anchorage-Dependent Mammalian Cells Growing in Suspension Bioreactors within a Three-Dimensional Hydrogel Microcarrier

doi: 10.3390/gels9030243

Figure Lengend Snippet: The production yield, activity, and purification of recombinant Cripto produced in the 3D microcarriers as compared to the 2D method. ( A ) SDS-PAGE analysis of the purification steps of recombinant Cripto: lane M is the protein molecular weight marker; lane 1 is the protein solution from ultrafiltration; lane 2 is the flowthrough after the first passage through the His-tag affinity Ni-NTA resin; lane 3 is the flowthrough after the second passage through the same resin; lane 4 is the first wash step; lane 5 is the second wash step; lane 6 is the first elution from the His-tag affinity Ni-NTA resin; and lane 7 is the second elution from the same resin. The band at approximately 27 kDa is the Cripto protein (indicated by the red arrow). ( B ) Quantitative amounts of Cripto protein produced in the 3D batch (Cripto (3D) ) with HEK293 cells encapsulated in PF microcarriers and incubated in bioreactors after three rounds of harvesting are compared to the maximum amount of Cripto produced in the 2D batch (Cripto (2D) ) with HEK293 cells adherent to cell culture plates and cultured to their density threshold limits. An initial cell seeding of 3.2 × 10 6 cells was used for both techniques. ( C ) The biological activity of recombinant Cripto was compared for Cripto produced in PF microcarriers versus the 2D method by measuring binding affinity to the AlK4 receptor. Four independent experiments were carried out for the 3D system and three independent experiments were performed for the 2D cultivation method. Results are shown as mean ± S.D. *** indicates p < 0.001.

Article Snippet: Serial dilutions of the concentrated protein were put into 96-well plates coated with either mouse Cripto antibody (for total concentration) or recombinant mouse activin receptor IB/Fc (for active protein concentration) (R&D systems AF1538 and 1477-AR, respectively).

Techniques: Activity Assay, Purification, Recombinant, Produced, SDS Page, Molecular Weight, Marker, Incubation, Cell Culture, Binding Assay

The effect of Cripto produced in 3D microcarriers on C2C12 cell proliferation detected by the BrdU incorporation assay. ( A ) C2C12 myoblast proliferation was evaluated by the BrdU incorporation assay after being cultured for 48 h in serum-free medium containing Cripto produced in 3D microcarriers (Cripto (3D) ) or commercially available Cripto (Cripto (R&D) ). Also evaluated were a bFGF medium positive control and a serum-free medium negative control. ( B ) The recombinant Cripto induces myoblast proliferation in a dose-dependent pattern; increasing concentrations of Cripto (3D) and Cripto (R&D) were added to C2C12 cells, and proliferation was quantified by the BrdU incorporation assay. ( C ) Cell proliferation was further evaluated by counting the total number of live cells. The proliferative effect of Cripto (3D) was compared with that of commercial Cripto (R&D) . The data are presented as mean ± S.D. from at least three independent experiments. * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

Journal: Gels

Article Title: Biomanufacturing Recombinantly Expressed Cripto-1 Protein in Anchorage-Dependent Mammalian Cells Growing in Suspension Bioreactors within a Three-Dimensional Hydrogel Microcarrier

doi: 10.3390/gels9030243

Figure Lengend Snippet: The effect of Cripto produced in 3D microcarriers on C2C12 cell proliferation detected by the BrdU incorporation assay. ( A ) C2C12 myoblast proliferation was evaluated by the BrdU incorporation assay after being cultured for 48 h in serum-free medium containing Cripto produced in 3D microcarriers (Cripto (3D) ) or commercially available Cripto (Cripto (R&D) ). Also evaluated were a bFGF medium positive control and a serum-free medium negative control. ( B ) The recombinant Cripto induces myoblast proliferation in a dose-dependent pattern; increasing concentrations of Cripto (3D) and Cripto (R&D) were added to C2C12 cells, and proliferation was quantified by the BrdU incorporation assay. ( C ) Cell proliferation was further evaluated by counting the total number of live cells. The proliferative effect of Cripto (3D) was compared with that of commercial Cripto (R&D) . The data are presented as mean ± S.D. from at least three independent experiments. * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

Article Snippet: Serial dilutions of the concentrated protein were put into 96-well plates coated with either mouse Cripto antibody (for total concentration) or recombinant mouse activin receptor IB/Fc (for active protein concentration) (R&D systems AF1538 and 1477-AR, respectively).

Techniques: Produced, BrdU Incorporation Assay, Cell Culture, Positive Control, Negative Control, Recombinant

FIGURE 1. Characterization of the recom- binant CR1 CCP 22–30_His fragment. (A) SDS-PAGE analysis of 4 mg CR1 CCP22-30 under reducing (R) and nonreducing (NR) conditions. The positions of the m.w. mark- ers are indicated. (B) CD spectroscopy of CR1 CCP22–30. A spectrum was recorded in the far-UV (200–260 nm) and collected six times. The mean values for each wavelength were calculated. The maximal ellipticity is indicated by an arrowhead. (C) Electron microscopy analysis of CR1 CCP22–30 (375000) after negative staining with 2% sodium silicotungstate.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Deciphering complement receptor type 1 interactions with recognition proteins of the lectin complement pathway.

doi: 10.4049/jimmunol.1202451

Figure Lengend Snippet: FIGURE 1. Characterization of the recom- binant CR1 CCP 22–30_His fragment. (A) SDS-PAGE analysis of 4 mg CR1 CCP22-30 under reducing (R) and nonreducing (NR) conditions. The positions of the m.w. mark- ers are indicated. (B) CD spectroscopy of CR1 CCP22–30. A spectrum was recorded in the far-UV (200–260 nm) and collected six times. The mean values for each wavelength were calculated. The maximal ellipticity is indicated by an arrowhead. (C) Electron microscopy analysis of CR1 CCP22–30 (375000) after negative staining with 2% sodium silicotungstate.

Article Snippet: Recombinant soluble human CR1 was purchased from R&D Systems Europe (Lille, France).

Techniques: SDS Page, Circular Dichroism, Electron Microscopy, Negative Staining

FIGURE 1. Lipid raft localization and release of CR-1 protein. A, NTERA2/D1 cells were stained with anti-CR-1 mAb (green), and lipid raft marker GM-1 was labeled with CTxB (red) without permeabilization and analyzed by confocal microscopy (upper panel). Magnified images for the marked regions in the upper panel are shown in the lower left panel. Negative controls without pri- mary antibody are shown in the lower right panel. Nuclei were stained with DAPI (blue) in all merged images. Scale bar 10 m. B, biochemical isolation of DRMs with 1% Triton X-100 by sucrose-gradient centrifugation of NTERA2/D1 cells. The gradient fractions (lanes 4–12) were analyzed with Western blot analysis for CR-1. Transferrin receptor (TfR) was used as a control for non-raft membrane protein. The lipid raft marker GM-1 in the same sam- ples was detected with HRP-conjugated CTxB by dot-blot analysis. C, compar- ative study of cell-associated (CL) and released (CM) CR-1 with or without serum by semi-quantitative Western blot analysis (see Table 1 and supple- mental Fig. S1, A–C). A representative blot for NTERA2/D1 cells is shown. The indicated amount of recombinant human CR-1 protein was used as a standard.

Journal: Journal of Biological Chemistry

Article Title: Growth Factor Induction of Cripto-1 Shedding by Glycosylphosphatidylinositol-Phospholipase D and Enhancement of Endothelial Cell Migration

doi: 10.1074/jbc.m702713200

Figure Lengend Snippet: FIGURE 1. Lipid raft localization and release of CR-1 protein. A, NTERA2/D1 cells were stained with anti-CR-1 mAb (green), and lipid raft marker GM-1 was labeled with CTxB (red) without permeabilization and analyzed by confocal microscopy (upper panel). Magnified images for the marked regions in the upper panel are shown in the lower left panel. Negative controls without pri- mary antibody are shown in the lower right panel. Nuclei were stained with DAPI (blue) in all merged images. Scale bar 10 m. B, biochemical isolation of DRMs with 1% Triton X-100 by sucrose-gradient centrifugation of NTERA2/D1 cells. The gradient fractions (lanes 4–12) were analyzed with Western blot analysis for CR-1. Transferrin receptor (TfR) was used as a control for non-raft membrane protein. The lipid raft marker GM-1 in the same sam- ples was detected with HRP-conjugated CTxB by dot-blot analysis. C, compar- ative study of cell-associated (CL) and released (CM) CR-1 with or without serum by semi-quantitative Western blot analysis (see Table 1 and supple- mental Fig. S1, A–C). A representative blot for NTERA2/D1 cells is shown. The indicated amount of recombinant human CR-1 protein was used as a standard.

Article Snippet: Reagents—Human CR-1 monoclonal antibodies (mAbs) (MAB2771 and FAB2772P) were obtained from R&D Systems (Minneapolis, MN) or developed as previously reported (B3F6) (8).

Techniques: Staining, Marker, Labeling, Confocal Microscopy, Isolation, Gradient Centrifugation, Western Blot, Control, Membrane, Dot Blot, Recombinant

Figure 1. CRIPTO micro-heterogeneity in ASC population (A) Schematic representation of the experimental design. (B) Representative FACS plots showing the gating strategy. The percentage of CRIPTO-positive cells above the threshold is indicated (bottom panel).

Journal: Developmental cell

Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.

doi: 10.1016/j.devcel.2023.11.009

Figure Lengend Snippet: Figure 1. CRIPTO micro-heterogeneity in ASC population (A) Schematic representation of the experimental design. (B) Representative FACS plots showing the gating strategy. The percentage of CRIPTO-positive cells above the threshold is indicated (bottom panel).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse Cripto Allophycocyanin MAb Biotechne - R&D systems Cat# FAB1538A Cripto Mouse specific Cell Signaling Technology Cat# 2818; RRID: AB_2240508 Anti-P4HB antibody [RL90] abcam Cat# ab2792; RRID: AB_303304 Pax7 Antibody Developmental Studies Hybridoma Bank (DHSB) Cat# PAX7; RRID: AB_528428 Anti-MyoD (C-20) Santa Cruz Biotechnology, Inc Cat# sc-304; RRID: AB_631992 Anti-GFP antibody abcam Cat# ab13970; RRID: AB_300798 Anti-human Ki67 Monosan Cat# MONX10283 Myogenin Antibody (F5D) Developmental Studies Hybridoma Bank (DHSB) Cat# F5D, RRID: AB_2146602 BrdU antibody [BU1/75 (ICR1)] GeneTex Cat# GTX26326; RRID: AB_1081056 Ki-67 Recombinant Rabbit Monoclonal Antibody (SP6) ThermoFisher Scientific Cat# MA5-14520; RRID: AB_10979488 Anti-Laminin Sigma-Aldrich L9393; RRID: AB_477163 Biotin-SP (long spacer) AffiniPure Goat Anti-Mouse IgG, Fcg subclass 1 specific Jackson ImmunoResearch Cat# 115-065-205, RRID: AB_2338571 Anti-mouse Alexa Fluor 549 ThermoFisher Scientific Cat# A-21203; RRID: AB_2535789 Anti-rabbit Alexa Fluor 488 ThermoFisher Scientific Cat# A-11001; RRID: AB_2534069 Anti-rabbit Alexa Fluor 555 ThermoFisher Scientific Cat# A-31572; RRID: AB_162543 Anti-rabbit Alexa Fluor 549 ThermoFisher Scientific Cat# A-21207; RRID: AB_141637 Alexa Fluor 488 AffiniPure Donkey Anti-Chicken IgY (IgG) (H+L) ThermoFisher Scientific Cat# 703-545-155, RRID: AB_2340375 Cy 3 AffiniPure Donkey Anti-Rat IgG (H+L) Jackson ImmunoResearch Cat# 712-165-153, RRID: AB_2340667 Chemicals, peptides, and recombinant proteins Sambucus Nigra Lectin (SNA, EBL), Biotinylated Vector Laboratories Cat# B-1305-2 Rhodamine Phalloidin ThermoFisher Scientific Cat# R415 Streptavidin-Cy3 conjugated Jackson ImmunoResearch Cat# 016-160-084 Streptavidin-488 conjugated Invitrogen Cat# S32354 Streptavidin-647 conjugated Invitrogen Cat# 532357 Tamoxifene Sigma-Aldrich Cat# T5648 Cardiotoxin Latoxan Laboratory Cat# L8102 Dispase II Roche Cat# 10887800 Collagenase A Roche Cat# 10103586001 DNAse I Roche Cat# 11284932001 Hank’s Balanced Salt Solution (HBSS) Gibco Cat# 14170-088 Bovine Serum Albumin (BSA) Sigma-Aldrich Cat# A4503 Penicillin-Streptomycin Gibco Cat# 15140-122 Collagenase Sigma-Aldrich Cat# C0130 Dulbecco’s Modified Eagle’s medium (DMEM) Gibco Cat# 41965-039 Ham’s F-12 Nutrient Mix, GlutaMAX Gibco Cat# 31765-027 Pyruvate Sodium Gibco Cat# 11360-039 L-Glutamine 200mM (100x) Gibco Cat# 25030-024 (Continued on next page) e1 Developmental Cell 58, 2896–2913.e1–e6, December 18, 2023

Techniques:

Figure 2. Transcriptional profiles of CRIPTONeg and CRIPTOPos cells and correlation between CRIPTO and PAX7 expression levels (A) Principal-component analysis (PCA) of CRIPTONeg and CRIPTOPos cells. Each dot represents individual mice. (B) Heatmap of the differential gene expression genes from individual mice (M1–M4). The color scale represents the normalized expression values of each gene.

Journal: Developmental cell

Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.

doi: 10.1016/j.devcel.2023.11.009

Figure Lengend Snippet: Figure 2. Transcriptional profiles of CRIPTONeg and CRIPTOPos cells and correlation between CRIPTO and PAX7 expression levels (A) Principal-component analysis (PCA) of CRIPTONeg and CRIPTOPos cells. Each dot represents individual mice. (B) Heatmap of the differential gene expression genes from individual mice (M1–M4). The color scale represents the normalized expression values of each gene.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse Cripto Allophycocyanin MAb Biotechne - R&D systems Cat# FAB1538A Cripto Mouse specific Cell Signaling Technology Cat# 2818; RRID: AB_2240508 Anti-P4HB antibody [RL90] abcam Cat# ab2792; RRID: AB_303304 Pax7 Antibody Developmental Studies Hybridoma Bank (DHSB) Cat# PAX7; RRID: AB_528428 Anti-MyoD (C-20) Santa Cruz Biotechnology, Inc Cat# sc-304; RRID: AB_631992 Anti-GFP antibody abcam Cat# ab13970; RRID: AB_300798 Anti-human Ki67 Monosan Cat# MONX10283 Myogenin Antibody (F5D) Developmental Studies Hybridoma Bank (DHSB) Cat# F5D, RRID: AB_2146602 BrdU antibody [BU1/75 (ICR1)] GeneTex Cat# GTX26326; RRID: AB_1081056 Ki-67 Recombinant Rabbit Monoclonal Antibody (SP6) ThermoFisher Scientific Cat# MA5-14520; RRID: AB_10979488 Anti-Laminin Sigma-Aldrich L9393; RRID: AB_477163 Biotin-SP (long spacer) AffiniPure Goat Anti-Mouse IgG, Fcg subclass 1 specific Jackson ImmunoResearch Cat# 115-065-205, RRID: AB_2338571 Anti-mouse Alexa Fluor 549 ThermoFisher Scientific Cat# A-21203; RRID: AB_2535789 Anti-rabbit Alexa Fluor 488 ThermoFisher Scientific Cat# A-11001; RRID: AB_2534069 Anti-rabbit Alexa Fluor 555 ThermoFisher Scientific Cat# A-31572; RRID: AB_162543 Anti-rabbit Alexa Fluor 549 ThermoFisher Scientific Cat# A-21207; RRID: AB_141637 Alexa Fluor 488 AffiniPure Donkey Anti-Chicken IgY (IgG) (H+L) ThermoFisher Scientific Cat# 703-545-155, RRID: AB_2340375 Cy 3 AffiniPure Donkey Anti-Rat IgG (H+L) Jackson ImmunoResearch Cat# 712-165-153, RRID: AB_2340667 Chemicals, peptides, and recombinant proteins Sambucus Nigra Lectin (SNA, EBL), Biotinylated Vector Laboratories Cat# B-1305-2 Rhodamine Phalloidin ThermoFisher Scientific Cat# R415 Streptavidin-Cy3 conjugated Jackson ImmunoResearch Cat# 016-160-084 Streptavidin-488 conjugated Invitrogen Cat# S32354 Streptavidin-647 conjugated Invitrogen Cat# 532357 Tamoxifene Sigma-Aldrich Cat# T5648 Cardiotoxin Latoxan Laboratory Cat# L8102 Dispase II Roche Cat# 10887800 Collagenase A Roche Cat# 10103586001 DNAse I Roche Cat# 11284932001 Hank’s Balanced Salt Solution (HBSS) Gibco Cat# 14170-088 Bovine Serum Albumin (BSA) Sigma-Aldrich Cat# A4503 Penicillin-Streptomycin Gibco Cat# 15140-122 Collagenase Sigma-Aldrich Cat# C0130 Dulbecco’s Modified Eagle’s medium (DMEM) Gibco Cat# 41965-039 Ham’s F-12 Nutrient Mix, GlutaMAX Gibco Cat# 31765-027 Pyruvate Sodium Gibco Cat# 11360-039 L-Glutamine 200mM (100x) Gibco Cat# 25030-024 (Continued on next page) e1 Developmental Cell 58, 2896–2913.e1–e6, December 18, 2023

Techniques: Expressing, Gene Expression

Figure 4. Regenerative potential of CRIPTONeg and CRIPTOPos cells and reconstitution of naı¨ve CRIPTO micro-heterogeneity following in vivo cell transplantation (A) Schematic representation of the experimental design. (B) Representative images of mosaic recipient muscles showing tdTomato-positive myofibers (RFP-red) and double staining with GFP (green) and LAMININ (white). (C and D) Quantification of tdTomato+ myofibers number/area (C) and GFP+ cells from CRIPTONeg and CRIPTOPos donor cells normalized per mm2 (D). Scale bars: 100 mm. Data are mean ± SEM. Nuclei were counter-stained with DAPI. (n = 3). (E) Schematic representation of the experimental design. (F and G) Representative dot plots of surface CRIPTO in GFP-positive CRIPTONeg and CRIPTOPos cells used for transplantation (F) and GFP-positive ASCs (3 dpi) derived from CRIPTONeg and CRIPTOPos -transplanted mice (G). The percentage of CRIPTO-positive cells above the threshold is indicated in the dot plots.

Journal: Developmental cell

Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.

doi: 10.1016/j.devcel.2023.11.009

Figure Lengend Snippet: Figure 4. Regenerative potential of CRIPTONeg and CRIPTOPos cells and reconstitution of naı¨ve CRIPTO micro-heterogeneity following in vivo cell transplantation (A) Schematic representation of the experimental design. (B) Representative images of mosaic recipient muscles showing tdTomato-positive myofibers (RFP-red) and double staining with GFP (green) and LAMININ (white). (C and D) Quantification of tdTomato+ myofibers number/area (C) and GFP+ cells from CRIPTONeg and CRIPTOPos donor cells normalized per mm2 (D). Scale bars: 100 mm. Data are mean ± SEM. Nuclei were counter-stained with DAPI. (n = 3). (E) Schematic representation of the experimental design. (F and G) Representative dot plots of surface CRIPTO in GFP-positive CRIPTONeg and CRIPTOPos cells used for transplantation (F) and GFP-positive ASCs (3 dpi) derived from CRIPTONeg and CRIPTOPos -transplanted mice (G). The percentage of CRIPTO-positive cells above the threshold is indicated in the dot plots.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse Cripto Allophycocyanin MAb Biotechne - R&D systems Cat# FAB1538A Cripto Mouse specific Cell Signaling Technology Cat# 2818; RRID: AB_2240508 Anti-P4HB antibody [RL90] abcam Cat# ab2792; RRID: AB_303304 Pax7 Antibody Developmental Studies Hybridoma Bank (DHSB) Cat# PAX7; RRID: AB_528428 Anti-MyoD (C-20) Santa Cruz Biotechnology, Inc Cat# sc-304; RRID: AB_631992 Anti-GFP antibody abcam Cat# ab13970; RRID: AB_300798 Anti-human Ki67 Monosan Cat# MONX10283 Myogenin Antibody (F5D) Developmental Studies Hybridoma Bank (DHSB) Cat# F5D, RRID: AB_2146602 BrdU antibody [BU1/75 (ICR1)] GeneTex Cat# GTX26326; RRID: AB_1081056 Ki-67 Recombinant Rabbit Monoclonal Antibody (SP6) ThermoFisher Scientific Cat# MA5-14520; RRID: AB_10979488 Anti-Laminin Sigma-Aldrich L9393; RRID: AB_477163 Biotin-SP (long spacer) AffiniPure Goat Anti-Mouse IgG, Fcg subclass 1 specific Jackson ImmunoResearch Cat# 115-065-205, RRID: AB_2338571 Anti-mouse Alexa Fluor 549 ThermoFisher Scientific Cat# A-21203; RRID: AB_2535789 Anti-rabbit Alexa Fluor 488 ThermoFisher Scientific Cat# A-11001; RRID: AB_2534069 Anti-rabbit Alexa Fluor 555 ThermoFisher Scientific Cat# A-31572; RRID: AB_162543 Anti-rabbit Alexa Fluor 549 ThermoFisher Scientific Cat# A-21207; RRID: AB_141637 Alexa Fluor 488 AffiniPure Donkey Anti-Chicken IgY (IgG) (H+L) ThermoFisher Scientific Cat# 703-545-155, RRID: AB_2340375 Cy 3 AffiniPure Donkey Anti-Rat IgG (H+L) Jackson ImmunoResearch Cat# 712-165-153, RRID: AB_2340667 Chemicals, peptides, and recombinant proteins Sambucus Nigra Lectin (SNA, EBL), Biotinylated Vector Laboratories Cat# B-1305-2 Rhodamine Phalloidin ThermoFisher Scientific Cat# R415 Streptavidin-Cy3 conjugated Jackson ImmunoResearch Cat# 016-160-084 Streptavidin-488 conjugated Invitrogen Cat# S32354 Streptavidin-647 conjugated Invitrogen Cat# 532357 Tamoxifene Sigma-Aldrich Cat# T5648 Cardiotoxin Latoxan Laboratory Cat# L8102 Dispase II Roche Cat# 10887800 Collagenase A Roche Cat# 10103586001 DNAse I Roche Cat# 11284932001 Hank’s Balanced Salt Solution (HBSS) Gibco Cat# 14170-088 Bovine Serum Albumin (BSA) Sigma-Aldrich Cat# A4503 Penicillin-Streptomycin Gibco Cat# 15140-122 Collagenase Sigma-Aldrich Cat# C0130 Dulbecco’s Modified Eagle’s medium (DMEM) Gibco Cat# 41965-039 Ham’s F-12 Nutrient Mix, GlutaMAX Gibco Cat# 31765-027 Pyruvate Sodium Gibco Cat# 11360-039 L-Glutamine 200mM (100x) Gibco Cat# 25030-024 (Continued on next page) e1 Developmental Cell 58, 2896–2913.e1–e6, December 18, 2023

Techniques: In Vivo, Transplantation Assay, Muscles, Double Staining, Staining, Derivative Assay

Figure 5. Cellular mechanisms of CRIPTO micro-heterogeneity reconstitution (A) Schematic representation of the experimental design. GFP-positive CRIPTONeg and CRIPTOPos ASCs were incubated at RT ± aCRIPTO-APC antibody (line a) or ± U73122 (line b) and stained with aCRIPTO-APC.

Journal: Developmental cell

Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.

doi: 10.1016/j.devcel.2023.11.009

Figure Lengend Snippet: Figure 5. Cellular mechanisms of CRIPTO micro-heterogeneity reconstitution (A) Schematic representation of the experimental design. GFP-positive CRIPTONeg and CRIPTOPos ASCs were incubated at RT ± aCRIPTO-APC antibody (line a) or ± U73122 (line b) and stained with aCRIPTO-APC.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse Cripto Allophycocyanin MAb Biotechne - R&D systems Cat# FAB1538A Cripto Mouse specific Cell Signaling Technology Cat# 2818; RRID: AB_2240508 Anti-P4HB antibody [RL90] abcam Cat# ab2792; RRID: AB_303304 Pax7 Antibody Developmental Studies Hybridoma Bank (DHSB) Cat# PAX7; RRID: AB_528428 Anti-MyoD (C-20) Santa Cruz Biotechnology, Inc Cat# sc-304; RRID: AB_631992 Anti-GFP antibody abcam Cat# ab13970; RRID: AB_300798 Anti-human Ki67 Monosan Cat# MONX10283 Myogenin Antibody (F5D) Developmental Studies Hybridoma Bank (DHSB) Cat# F5D, RRID: AB_2146602 BrdU antibody [BU1/75 (ICR1)] GeneTex Cat# GTX26326; RRID: AB_1081056 Ki-67 Recombinant Rabbit Monoclonal Antibody (SP6) ThermoFisher Scientific Cat# MA5-14520; RRID: AB_10979488 Anti-Laminin Sigma-Aldrich L9393; RRID: AB_477163 Biotin-SP (long spacer) AffiniPure Goat Anti-Mouse IgG, Fcg subclass 1 specific Jackson ImmunoResearch Cat# 115-065-205, RRID: AB_2338571 Anti-mouse Alexa Fluor 549 ThermoFisher Scientific Cat# A-21203; RRID: AB_2535789 Anti-rabbit Alexa Fluor 488 ThermoFisher Scientific Cat# A-11001; RRID: AB_2534069 Anti-rabbit Alexa Fluor 555 ThermoFisher Scientific Cat# A-31572; RRID: AB_162543 Anti-rabbit Alexa Fluor 549 ThermoFisher Scientific Cat# A-21207; RRID: AB_141637 Alexa Fluor 488 AffiniPure Donkey Anti-Chicken IgY (IgG) (H+L) ThermoFisher Scientific Cat# 703-545-155, RRID: AB_2340375 Cy 3 AffiniPure Donkey Anti-Rat IgG (H+L) Jackson ImmunoResearch Cat# 712-165-153, RRID: AB_2340667 Chemicals, peptides, and recombinant proteins Sambucus Nigra Lectin (SNA, EBL), Biotinylated Vector Laboratories Cat# B-1305-2 Rhodamine Phalloidin ThermoFisher Scientific Cat# R415 Streptavidin-Cy3 conjugated Jackson ImmunoResearch Cat# 016-160-084 Streptavidin-488 conjugated Invitrogen Cat# S32354 Streptavidin-647 conjugated Invitrogen Cat# 532357 Tamoxifene Sigma-Aldrich Cat# T5648 Cardiotoxin Latoxan Laboratory Cat# L8102 Dispase II Roche Cat# 10887800 Collagenase A Roche Cat# 10103586001 DNAse I Roche Cat# 11284932001 Hank’s Balanced Salt Solution (HBSS) Gibco Cat# 14170-088 Bovine Serum Albumin (BSA) Sigma-Aldrich Cat# A4503 Penicillin-Streptomycin Gibco Cat# 15140-122 Collagenase Sigma-Aldrich Cat# C0130 Dulbecco’s Modified Eagle’s medium (DMEM) Gibco Cat# 41965-039 Ham’s F-12 Nutrient Mix, GlutaMAX Gibco Cat# 31765-027 Pyruvate Sodium Gibco Cat# 11360-039 L-Glutamine 200mM (100x) Gibco Cat# 25030-024 (Continued on next page) e1 Developmental Cell 58, 2896–2913.e1–e6, December 18, 2023

Techniques: Incubation, Staining

Figure 7. CRIPTO micro-heterogeneity preserves the balance between self-renewal and myogenic commitment (A) Representative pictures of PAX7 (magenta) and GFP (green) staining (upper panel) and quantification of PAX7+/GFP± cells (bottom panel) in Criptowt and CriptocKO muscle sections. Inset shows higher magnification (103) of the boxed area. Scale bars: 75 mm.

Journal: Developmental cell

Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.

doi: 10.1016/j.devcel.2023.11.009

Figure Lengend Snippet: Figure 7. CRIPTO micro-heterogeneity preserves the balance between self-renewal and myogenic commitment (A) Representative pictures of PAX7 (magenta) and GFP (green) staining (upper panel) and quantification of PAX7+/GFP± cells (bottom panel) in Criptowt and CriptocKO muscle sections. Inset shows higher magnification (103) of the boxed area. Scale bars: 75 mm.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse Cripto Allophycocyanin MAb Biotechne - R&D systems Cat# FAB1538A Cripto Mouse specific Cell Signaling Technology Cat# 2818; RRID: AB_2240508 Anti-P4HB antibody [RL90] abcam Cat# ab2792; RRID: AB_303304 Pax7 Antibody Developmental Studies Hybridoma Bank (DHSB) Cat# PAX7; RRID: AB_528428 Anti-MyoD (C-20) Santa Cruz Biotechnology, Inc Cat# sc-304; RRID: AB_631992 Anti-GFP antibody abcam Cat# ab13970; RRID: AB_300798 Anti-human Ki67 Monosan Cat# MONX10283 Myogenin Antibody (F5D) Developmental Studies Hybridoma Bank (DHSB) Cat# F5D, RRID: AB_2146602 BrdU antibody [BU1/75 (ICR1)] GeneTex Cat# GTX26326; RRID: AB_1081056 Ki-67 Recombinant Rabbit Monoclonal Antibody (SP6) ThermoFisher Scientific Cat# MA5-14520; RRID: AB_10979488 Anti-Laminin Sigma-Aldrich L9393; RRID: AB_477163 Biotin-SP (long spacer) AffiniPure Goat Anti-Mouse IgG, Fcg subclass 1 specific Jackson ImmunoResearch Cat# 115-065-205, RRID: AB_2338571 Anti-mouse Alexa Fluor 549 ThermoFisher Scientific Cat# A-21203; RRID: AB_2535789 Anti-rabbit Alexa Fluor 488 ThermoFisher Scientific Cat# A-11001; RRID: AB_2534069 Anti-rabbit Alexa Fluor 555 ThermoFisher Scientific Cat# A-31572; RRID: AB_162543 Anti-rabbit Alexa Fluor 549 ThermoFisher Scientific Cat# A-21207; RRID: AB_141637 Alexa Fluor 488 AffiniPure Donkey Anti-Chicken IgY (IgG) (H+L) ThermoFisher Scientific Cat# 703-545-155, RRID: AB_2340375 Cy 3 AffiniPure Donkey Anti-Rat IgG (H+L) Jackson ImmunoResearch Cat# 712-165-153, RRID: AB_2340667 Chemicals, peptides, and recombinant proteins Sambucus Nigra Lectin (SNA, EBL), Biotinylated Vector Laboratories Cat# B-1305-2 Rhodamine Phalloidin ThermoFisher Scientific Cat# R415 Streptavidin-Cy3 conjugated Jackson ImmunoResearch Cat# 016-160-084 Streptavidin-488 conjugated Invitrogen Cat# S32354 Streptavidin-647 conjugated Invitrogen Cat# 532357 Tamoxifene Sigma-Aldrich Cat# T5648 Cardiotoxin Latoxan Laboratory Cat# L8102 Dispase II Roche Cat# 10887800 Collagenase A Roche Cat# 10103586001 DNAse I Roche Cat# 11284932001 Hank’s Balanced Salt Solution (HBSS) Gibco Cat# 14170-088 Bovine Serum Albumin (BSA) Sigma-Aldrich Cat# A4503 Penicillin-Streptomycin Gibco Cat# 15140-122 Collagenase Sigma-Aldrich Cat# C0130 Dulbecco’s Modified Eagle’s medium (DMEM) Gibco Cat# 41965-039 Ham’s F-12 Nutrient Mix, GlutaMAX Gibco Cat# 31765-027 Pyruvate Sodium Gibco Cat# 11360-039 L-Glutamine 200mM (100x) Gibco Cat# 25030-024 (Continued on next page) e1 Developmental Cell 58, 2896–2913.e1–e6, December 18, 2023

Techniques: Staining