Journal: Cell reports

Article Title: Tracheal separation is driven by NKX2-1-mediated repression of Efnb2 and regulation of endodermal cell sorting

doi: 10.1016/j.celrep.2022.110510

Figure Lengend Snippet:

Article Snippet: Immunofluorescence was performed according to standard techniques with antibodies against acetylated α-tubulin (1:250, Sigma, T7451), BrdU (1:150, Abcam, ab6326), cleaved caspase3 (1:200, Cell Signaling, 9661), EPHA4 (1:75, R&D, AF641), EPHB2 (1:75, R&D, AF467), EPHB3 (1:75, R&D, AF432), EPHB4 (1:75, R&D, AF446), phospho-EPHA2+A3+A4 (1:150, Abcam, ab62256), phospho-EPHB1+B2 (1:150, Abcam, ab61791), GFP (1:500, Abcam, ab13970), KRT5 (1:250, BioLegend, 905501), KRT8 (1:75, DSHB, AB_531826), LRIG1 (1:200, R&D, AF3688), MUC5B (1:300, Santa Cruz Biotechnology, sc-20119), NKX2-1 (1:150, Millipore, 07–601), NKX2-1 (1:100, Santa Cruz Biotechnology, sc-8761), NKX2-1 (1:300, ThermoFisher, MS-699), RFP (1:250, Abcam, ab62341), SOX2 (1:300, Neuromics, GT15098), and SOX2 (1:250, Seven Hills Bioreagents, WRAB-1236).

Techniques: Recombinant, Transfection, Multiplex Assay, Reporter Assay

Journal: eNeuro

Article Title: Common Origin of the Cerebellar Dual Somatotopic Areas Revealed by Tracking Embryonic Purkinje Cell Clusters with Birthdate Tagging

doi: 10.1523/ENEURO.0251-20.2020

Figure Lengend Snippet: Antibodies used in the study

Article Snippet: Goat anti-EphA4 (R&D Systems), goat anti-FoxP2 (Everest Biotech), rabbit anti-Corl2 (provided by Dr. Ono at KAN Research Institute) and rat anti-OL-protocadherin (Millipore) are the primary antibodies used in the majority of experiments.

Techniques: Concentration Assay, Affinity Purification

Journal: eNeuro

Article Title: Common Origin of the Cerebellar Dual Somatotopic Areas Revealed by Tracking Embryonic Purkinje Cell Clusters with Birthdate Tagging

doi: 10.1523/ENEURO.0251-20.2020

Figure Lengend Snippet: Purkinje cell clusters in coronal sections of the left E16.5 mouse cerebellum identified by marker expression profiles. A–I , Sections at different caudorostral levels indicated by percentile. Fluorescent signals of immunostaining for EphA4 (red), Pcdh10 (blue) and FoxP2 (green) are merged. White dashed lines indicate the boundary of recognized clusters. Inset in each panel shows drawings of recognized clusters at half magnification. Names of each cluster were given later based on the lineage analysis (c.f. ). J , Dorsal view of the 3D reconstruction of clusters. Red lines indicate the rostrocaudal level of the section in each panel. Scale bar in A (100 μm) applies to panels A–I . Abbreviations, c-l, c-m, d, dl, l, m, ml, rdl, vl, names of the lineage of clusters; C, caudal; D, dorsal; DCN, deep cerebellar nucleus; L, lateral; M, medial; R, rostral.V, ventral.

Article Snippet: Goat anti-EphA4 (R&D Systems), goat anti-FoxP2 (Everest Biotech), rabbit anti-Corl2 (provided by Dr. Ono at KAN Research Institute) and rat anti-OL-protocadherin (Millipore) are the primary antibodies used in the majority of experiments.

Techniques: Marker, Expressing, Immunostaining

Journal: eNeuro

Article Title: Common Origin of the Cerebellar Dual Somatotopic Areas Revealed by Tracking Embryonic Purkinje Cell Clusters with Birthdate Tagging

doi: 10.1523/ENEURO.0251-20.2020

Figure Lengend Snippet: Purkinje cell clusters in coronal sections of the left E17.5 mouse cerebellum identified by marker expression profiles. A–I , Sections at different caudorostral levels indicated by percentile. Fluorescent signals of immunostaining for EphA4 (red), Pcdh10 (blue) and FoxP2 (green) are merged. White dashed lines indicate the boundary of recognized clusters. Inset in each panel shows drawings of recognized clusters at half magnification. Names of each cluster were basically adopted from , but see Results. J , Dorsal view of the 3D reconstruction of clusters. Yellow lines indicate immature fissures. Red lines indicate the rostrocaudal level of the section in each panel. Scale bar in A (100 μm) applies to panels A–I . Abbreviations, fl1-2, fl4, ha2-3, ha4, ha6, hc1, hc2, hp1-2, hp4, ia1-2, ia3-5, ia4, ic1-2, ic3, ip1-2, it2, it3, no1-2, no3, no4, pf1-2, va1, va2-4, vc1, vp1-2, vp3-4, vt1, vt2, vt3, vt4, vt5, names of E17.5 clusters; C, caudal; D, dorsal; DCN, deep cerebellar nucleus; L, lateral; M, medial; R, rostral.V, ventral.

Article Snippet: Goat anti-EphA4 (R&D Systems), goat anti-FoxP2 (Everest Biotech), rabbit anti-Corl2 (provided by Dr. Ono at KAN Research Institute) and rat anti-OL-protocadherin (Millipore) are the primary antibodies used in the majority of experiments.

Techniques: Marker, Expressing, Immunostaining

Journal: eNeuro

Article Title: Common Origin of the Cerebellar Dual Somatotopic Areas Revealed by Tracking Embryonic Purkinje Cell Clusters with Birthdate Tagging

doi: 10.1523/ENEURO.0251-20.2020

Figure Lengend Snippet: Definition of E17.5 PC clusters

Article Snippet: Goat anti-EphA4 (R&D Systems), goat anti-FoxP2 (Everest Biotech), rabbit anti-Corl2 (provided by Dr. Ono at KAN Research Institute) and rat anti-OL-protocadherin (Millipore) are the primary antibodies used in the majority of experiments.

Techniques:

Journal: eNeuro

Article Title: Common Origin of the Cerebellar Dual Somatotopic Areas Revealed by Tracking Embryonic Purkinje Cell Clusters with Birthdate Tagging

doi: 10.1523/ENEURO.0251-20.2020

Figure Lengend Snippet: Nine E14.5 PC clusters, their molecular expression profiles, PC birthdates and fates at E17.5

Article Snippet: Goat anti-EphA4 (R&D Systems), goat anti-FoxP2 (Everest Biotech), rabbit anti-Corl2 (provided by Dr. Ono at KAN Research Institute) and rat anti-OL-protocadherin (Millipore) are the primary antibodies used in the majority of experiments.

Techniques: Expressing

Journal: eNeuro

Article Title: Common Origin of the Cerebellar Dual Somatotopic Areas Revealed by Tracking Embryonic Purkinje Cell Clusters with Birthdate Tagging

doi: 10.1523/ENEURO.0251-20.2020

Figure Lengend Snippet: Spatial distribution of E10.5-, E11.5-, and E12.5-born PCs in the E14.5 cerebellum. A–D , Images of coronal sections of the left cerebellum of G2A::Ai9 embryos which had tamoxifen injection at E10.5 (subpanel 1), E11.5 (subpanel 2) and E12.5 (subpanel 3) at different rostrocaudal levels. The top, center and bottom panels show images of the tdTomato fluorescence signal (top), three signals merged, and signals of tdTomato fluorescence (green) and immunostaining signals of Pcd10 (red) and EphA4 (blue) in ( A ), while only the tdTomato fluorescence signal is shown in B–D . The tdTomato signal indicates neurons (mostly PCs) that expressed Neurog2 -CreER at the time of tamoxifen injection. White dashed lines indicate the coutour of the cerebellum. Subpanel 4 shows schematic drawings of identified clusters. Blue and Orange asterisks indicate (position of) the E10.5-PC-sparse and E12.5-PC-sparse clusters, respectively. E , Relative labeling density in nine E14.5 clusters. The plotted data were the average of the tdTomato fluorescence signal in the digital file (0-255) measured in 9-18 square areas of 900 μm 2 located inside the identified cluster in 3-6 sections. The same brightness and contrast adjustment was done in all sections. F , Confocal high-magnification image of immunostaining and tdTomato expression at the junction between the ml and c-l clusters in the AM10.5 G2A::Ai9 embryo, showing expression of tdTomato (E10.5-born neurons) colocalized exactly with Corl2 (PC marker) at the cellular level in the ml cluster. The area of this image is indicated in B4 with a square. Abbreviations, c-l, c-m, d, dl, l, m, ml, rdl, vl, names of E14.5 clusters; C, caudal; D, dorsal; L, lateral; M, medial; R, rostral.V, ventral.

Article Snippet: Goat anti-EphA4 (R&D Systems), goat anti-FoxP2 (Everest Biotech), rabbit anti-Corl2 (provided by Dr. Ono at KAN Research Institute) and rat anti-OL-protocadherin (Millipore) are the primary antibodies used in the majority of experiments.

Techniques: Injection, Fluorescence, Immunostaining, Labeling, Expressing, Marker

Journal: eNeuro

Article Title: Common Origin of the Cerebellar Dual Somatotopic Areas Revealed by Tracking Embryonic Purkinje Cell Clusters with Birthdate Tagging

doi: 10.1523/ENEURO.0251-20.2020

Figure Lengend Snippet: Identification of E10.5-PC-sparse and E12.5-PC-sparse clusters in the E14.5-E17.5 cerebellums. A–D , Images of coronal sections of the left TM10.5 and TM12.5 cerebellums at nearly the same level (subpanel 1 and 2, respectively) and schematic drawing of clusters (subpanel 3) and dorsal view of the 3D scheme (subpanel 4) of the E14.5 ( A ), E15.5 ( B ), E16.5 ( C ), and E17.5 ( D ) cerebellums. In subpanels 1 and 2, the tdTomato fluorescence signal (top), three signals merged, and signals of tdTomato fluorescence (green) and immunostaining signals of Pcd10 (red) and EphA4 (blue) are shown from the top to the bottom. Blue and orange colors in dashed lines (in subpanels in 1 and 2), circumscribing lines (in subpanels 3), and clusters in the 3D schemes (in subpanels 4) indicate E10.5-PC-sparse and E12.5-PC-sparse clusters. Black line in the 3D scheme indicates the position of the coronal section in subpanels 1-3. Abbreviations, c-l, c-m, d, dl, l, m, ml, rdl, vl, names of E14.5 clusters; ha2-3, ha4, ha6, hc1, hc2, hp1-2, hp4, ia1-2, ia4, ic1-2, ic3, ip1-2, it2, it3, va1, va2-4, vc1, vp1-2, vp3-4, vt1, vt2, vt3, vt4, vt5, names of E17.5 clusters; C, caudal; D, dorsal; L, lateral; M, medial; R, rostral.V, ventral.

Article Snippet: Goat anti-EphA4 (R&D Systems), goat anti-FoxP2 (Everest Biotech), rabbit anti-Corl2 (provided by Dr. Ono at KAN Research Institute) and rat anti-OL-protocadherin (Millipore) are the primary antibodies used in the majority of experiments.

Techniques: Fluorescence, Immunostaining

Journal: eNeuro

Article Title: Common Origin of the Cerebellar Dual Somatotopic Areas Revealed by Tracking Embryonic Purkinje Cell Clusters with Birthdate Tagging

doi: 10.1523/ENEURO.0251-20.2020

Figure Lengend Snippet: Changes in the molecular expression profile in PC clusters from E14.5 to E17.5

Article Snippet: Goat anti-EphA4 (R&D Systems), goat anti-FoxP2 (Everest Biotech), rabbit anti-Corl2 (provided by Dr. Ono at KAN Research Institute) and rat anti-OL-protocadherin (Millipore) are the primary antibodies used in the majority of experiments.

Techniques: Expressing

Journal: eNeuro

Article Title: Common Origin of the Cerebellar Dual Somatotopic Areas Revealed by Tracking Embryonic Purkinje Cell Clusters with Birthdate Tagging

doi: 10.1523/ENEURO.0251-20.2020

Figure Lengend Snippet: Separation and migration of the c-l lineage cluster from E14.5 to E17.5. A , Horizontal sections of the left paravermal and hemispheric cerebellum at around the central level. Sections were immunostained for Corl2 (green channel), EphA4 (blue channel), and Pcdh10 (red channel). Dashed lines circumscribe c-l lineage clusters. Arrowheads indicate the lateral migration of the rostral c-l lineage clusters. B , Dorsal view of the three-dimensional scheme of reconstructed c-l lineage clusters (blue). In the bottom panel, ml and c-m lineage clusters (orange and white) are added. Red transversal line indicate the position of the coronal section shown in ( B ). Black dashed line indicate the contour of the ia3-5 cluster which is located ventral to the it3 cluster in ( B4 ). C , Images of a part of the left cerebellum at the junction between the c-l and c-m lineage clusters in coronal sections. The top image shows merged signals of tdTomato expression, which indicates E10.5-born PCs (green) and immunostaining for EphA4 (blue) and Pcdh10 (red). The bottom subpanel shows only the image of only tdTomato labeling. D , Schematic drawing of clusters shown in C . In A–D , columns of subpanels 1-4 are from E14.5, E15.5, E16.5 and E17.5 cerebellums. Dashed lines indicate c-l lineage clusters that are E10.5-PC-sparse and weak in Pcdh10 expression in A and C . Single asterisks indicate the most medial part of the c-l cluster or the most medial c-l lineage cluster which had higher expression of Corl2 than the rest of the c-l lineage clusters in A–D . Double asterisks indicate the most lateral part of the c-m cluster or the most lateral c-m lineage cluster which intercalated the c-l lineage clusters in A–D . Arrows indicate the direction of cluster migration. Scale bar in C1 (100 μm) applies to C1–C4 . Abbreviations, c-l, c-m, d, ml, names of E14.5 clusters; ha4, ia1-2, ia3-5, ia4, ic1-2, ic3, ip1-2, it2, it3, va2-4, names of E17.5 clusters; C, caudal; D, dorsal; L, lateral; M, medial; R, rostral.V, ventral.

Article Snippet: Goat anti-EphA4 (R&D Systems), goat anti-FoxP2 (Everest Biotech), rabbit anti-Corl2 (provided by Dr. Ono at KAN Research Institute) and rat anti-OL-protocadherin (Millipore) are the primary antibodies used in the majority of experiments.

Techniques: Migration, Expressing, Immunostaining, Labeling