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Image Search Results
Journal: bioRxiv
Article Title: Epithelial Vegfa specifies a distinct endothelial population in the mouse lung
doi: 10.1101/840033
Figure Lengend Snippet: (A) tSNE plot of scRNA-seq results from sorted P14 lung ECs with each cell population color-coded and the corresponding cell number shown in parenthesis. Lower panels: gene expression showing that Car4 marks Car4 ECs; Plvap marks Plvap ECs, Vwf ECs, and lymphatic ECs. Epithelial and mesenchymal populations are minor contaminants from sorting. (B) Violin plots showing markers used to identify the six cell populations. (C) Heat map showing top 5 genes of each EC population. Plvap is expressed by all non-Car4 ECs, as shown in (A), and thus not among the top genes for Plvap ECs.
Article Snippet: The following antibodies were used: rabbit anti-Aquaporin 5 (AQP5, 1:2500, ab78486, Abcam),
Techniques: Expressing
Journal: bioRxiv
Article Title: Epithelial Vegfa specifies a distinct endothelial population in the mouse lung
doi: 10.1101/840033
Figure Lengend Snippet: (A) Representative en face view of immunostaining images from at least 5 mice. Boxed region is magnified as a section view in the first three images of the bottom row. CAR4 staining covers, whereas PLVAP staining surrounds, alveolar islands (dash). Perinuclear CAR4 and PLVAP staining allows assignment of ERG nuclei to CAR4 (cyan arrowhead) versus PLVAP (magenta arrowhead) ECs, which is automatically identified, as shown in the lower rightmost image (grey nuclei are ambiguous). Scale: 10 um. (B) Wholemount immunostaining of lungs with sparsely-labeled ECs, representative of at least 5 mice, viewed as a stack (40 um), a slab (top 20 um), or a section (1 um). Accumulation of tdT to ERG nuclei allows cell numeration (1 through 5). Cell #1 is a Car4 EC and cells #2-5 are non-Car4 ECs. Line profile analysis shows aligned versus shifted peaks for Car4 versus non-Car4 ECs, respectively. For shifted peaks, Car4 ECs are closer to the airspace than non-Car4 ECs (e.g. cell #3). Asterisk, avascular tissue surrounded by a single net-like Car4 EC. Open arrow, CDH5 junction overlapping with a single Car4 EC. Cell perimeter is measured by connecting protrusions that are visible in a projection view. Tam, 0.25 ug tamoxifen. Scale: 10 um. (C) Quantification of cell perimeter and comparison using Student’s t-test.
Article Snippet: The following antibodies were used: rabbit anti-Aquaporin 5 (AQP5, 1:2500, ab78486, Abcam),
Techniques: Immunostaining, Staining, Labeling
Journal: bioRxiv
Article Title: Epithelial Vegfa specifies a distinct endothelial population in the mouse lung
doi: 10.1101/840033
Figure Lengend Snippet: Wholemount immunostaining of lungs (A) and retinas (B) with sparsely-labeled ECs, representative of at least 5 mice. Tam, 0.25 ug tamoxifen. Scale: 10 um. (A) Cells #1-2 are Car4 ECs and cells #3-6 are non-Car4 ECs. ECs in non-capillary vessels (#4-6) are more elongated along the direction of blood flow. (B) Retinal macrophages (m), but not ECs, express CAR4. Tip ECs display characteristic filopodia (bracket) without a discernable lumen (weak ICAM2). Non-capillary ECs are similarly elongated, but none of the retinal ECs feature the net-like morphology of lung Car4 ECs.
Article Snippet: The following antibodies were used: rabbit anti-Aquaporin 5 (AQP5, 1:2500, ab78486, Abcam),
Techniques: Immunostaining, Labeling
Journal: bioRxiv
Article Title: Epithelial Vegfa specifies a distinct endothelial population in the mouse lung
doi: 10.1101/840033
Figure Lengend Snippet: (A) Violin plots of lung EC scRNA-seq data from showing retinal tip and stalk EC genes. Apln and Kdr are enriched in Car4 ECs; Aplnr , Tek (also known as Tie2 ), and Nrarp are enriched in Plvap ECs. (B) tSNE plot showing Esm1 expression in sporadic Plvap ECs (arrowhead in insert). (C) Dot plot of 25 recently identified tip EC genes with those enriched in Car4 ECs highlighted in red. (D) Representative wholemount immunostaining images of at least 2 mice. ESM1 is expressed by retinal tip ECs (filled arrowhead), as well as lung ECs near lobe edge in embryos (filled arrowhead), but in transitional ECs between capillaries and non-capillaries (open arrowhead). CAR4 expression initiates at E19 (open arrowhead), concomitant with AT1 cell differentiation. DLL4 staining is stronger in retinal arteries (a) than veins (v), distinguishable by vessel diameter, and is wide-spread and cord-like (dash) in the lung. Scale: 10 um.
Article Snippet: The following antibodies were used: rabbit anti-Aquaporin 5 (AQP5, 1:2500, ab78486, Abcam),
Techniques: Expressing, Immunostaining, Cell Differentiation, Staining
Journal: bioRxiv
Article Title: Epithelial Vegfa specifies a distinct endothelial population in the mouse lung
doi: 10.1101/840033
Figure Lengend Snippet: (A) Representative wholemount immunostaining images of at least 2 mice showing Apln CreER labels ECs but not specifically Car4 ECs. The specificity is quantified for 2 lungs and found to be variable. Filled arrowhead, CAR4 and GFP double-positive ECs; open arrowhead, GFP single positive ECs. Tam, 250 ug tamoxifen. Scale: 10 um. (B) Apln undergoes X-inactivation. FACS purification of P14 lung ECs (0.5 mg tamoxifen at P13) that are labeled (red box) or unlabeled (blue box) with native fluorescence from Rosa tdT , which are subjected to RT-PCR analysis. Labeled ECs, which are expected to express CreER, do not express Apln despite the presence of the wildtype allele, indicating X-inactivation. The same is true for P35 lung ECs (2 mg tamoxifen at P34). (C) tSNE plots of scRNA-seq results from sorted lung ECs from littermate Apln mutant and control males showing deletion of Apln but no change in the cell number and gene expression of Car4 and Plvap ECs. Asterisk, contaminating immune, epithelial, and mesenchymal cells (non-EC).
Article Snippet: The following antibodies were used: rabbit anti-Aquaporin 5 (AQP5, 1:2500, ab78486, Abcam),
Techniques: Immunostaining, Purification, Labeling, Fluorescence, Reverse Transcription Polymerase Chain Reaction, Mutagenesis, Expressing
Journal: bioRxiv
Article Title: Epithelial Vegfa specifies a distinct endothelial population in the mouse lung
doi: 10.1101/840033
Figure Lengend Snippet: (A) Left panels: tSNE plot of scRNA-seq results from sorted ECs from littermate lungs showing a specific, complete loss of Car4 ECs in the mutant with the percentages out of all ECs in parenthesis. Plvap ECs and proliferating ( Mki67 ) ECs are unaffected. Cell populations are color-coded as in . Mes, mesenchyme; Epi, epithelium; Lym, lymphatic EC; Vwf, Vwf EC. Compared to P14 , P7 lungs have more proliferating ( Mki67 ) ECs; their percentage is calculated with a UMI cutoff of 1. Right panels: en face view of immunostained littermate lungs, representative of at least 3 littermate pairs, showing rare Car4 staining in the remaining vessels in the mutant (open arrowhead). m, macrophage. Scale: 10 um. (B) Section immunostaining images of littermate lungs representative of at least 3 littermate pairs. As diagrammed, Car4 vessels (filled green arrowhead) abut the epithelium (AQP5), separated with a thin basement membrane (weak COL4 staining; dash open arrowhead) with no intervening pericytes (CSPG4; dash open arrowhead), but their sides away from the air space have a thicker basement membrane (strong COL4 staining; solid open arrowhead) and pericytes (solid open arrowhead). Non-Car4 vessels (filled white arrowhead; a subset of vessels in the control and all vessels in the mutant) do not abut the epithelium and are surrounded by a thick basement membrane and pericytes (solid open arrowhead). Scale: 10 um.
Article Snippet: The following antibodies were used: rabbit anti-Aquaporin 5 (AQP5, 1:2500, ab78486, Abcam),
Techniques: Mutagenesis, Staining, Immunostaining
Journal: bioRxiv
Article Title: Epithelial Vegfa specifies a distinct endothelial population in the mouse lung
doi: 10.1101/840033
Figure Lengend Snippet: (A) Left panels: en face view of immunostained littermate lungs, representative of at least 3 littermate pairs, showing a smoother surface (RAGE) of alveolar islands that are not subdivided by Car4 vessels in the mutant. Scale: 10 um. Right panels: H&E section images of littermate lungs with the corresponding mean linear intercept (MLI) and D 2 measurements. Scale: 100 um. Each symbol represents one mouse and is the average of 3 regions (882 um x 664 um; Student’s t-test). (B) En face view of immunostained littermate lungs, representative of at least 3 littermate pairs, showing fewer pericytes (PDGFRB; open arrowhead) but normal myofibroblasts (SMA/TAGLN/PDGFRA triple positive, although variable in staining intensity; filled arrowhead) in the mutant. Boxed regions are magnified in merged views showing costaining of SMA, TAGLN, and PDGFRA that is distinct from PDGFRB. Scale: 10 um. (C) Schematics with color-coded cell types showing AT1 derived Vegfa signals to Car4 ECs, which, together with myofibroblasts, promote secondary septation and persist in the resulting septae even after disappearance of myofibroblasts in the mature lung. The Vegfa mutant fails to form Car4 ECs; non-Car4 ECs and myofibroblasts are insufficient for secondary septation, resulting in alveolar enlargement. Note that Car4 vessels may consist of Car4 ECs and non-Car4 ECs, as diagrammed in . Alv, alveolus.
Article Snippet: The following antibodies were used: rabbit anti-Aquaporin 5 (AQP5, 1:2500, ab78486, Abcam),
Techniques: Mutagenesis, Staining, Derivative Assay