Journal: Immunity

Article Title: Gut commensals suppress epithelial cell retinoic acid synthesis to regulate intestinal interleukin-22 activity and prevent microbial dysbiosis.

doi: 10.1016/j.immuni.2018.11.018

Figure Lengend Snippet: Highlights

Article Snippet: All-trans-retinoic acid-d5 , Toronto Research Chemicals , Cat#: R250202.

Techniques: Cell Stimulation, Recombinant, Lysis, Reverse Transcription, Electron Microscopy, Single Photon Emission Computed Tomography, Staining, Isolation, Western Blot, Software, Sequencing

Journal: Developmental biology

Article Title: Cerebrovascular defects in Foxc1 mutants correlate with aberrant WNT and VEGF-A pathways downstream of retinoic acid from the meninges

doi: 10.1016/j.ydbio.2016.09.019

Figure Lengend Snippet: (A) Schematic depicting atRA diet exposure paradigm. (B) E14.5 non-treated and atRA treated control and Foxc1hith/lacz mutant whole embryos. Arrows indicate cerebral hemispheres. (C-J) Low magnification (C-F) images of GLUT1 immunohistochemistry on E14.5 coronal brain sections at the level of the forebrain. Higher magnification images (G-J) of GLUT1 in the neocortex highlight the morphology of the cerebrovasculature. Arrows indicated PNVP-derived vessels at their entry point in the cerebral wall. (K-M) Quantification of vessel branch points, number of blood vessels growing into the cortex from the PNVP and vessel diameter in untreated (n=3) and atRA treated control (n=3) and Foxc1hith/lacZ mutants (n=3). Asterisks indicate statistically significant difference (p<0.05) from non-atRA treated condition. Hashtag (#) indicates a statistically significant difference (p<0.05) from a non-atRA treated Foxc1 mutant. (N-Q) Immunohistochemistry of E14.5 cerebral cortex from non-treated (N, O) and atRA-treated (P, Q) wildtype and Foxc1hith/lacz mutant labeled with endothelial cell nuclear marker ERG1 and BrdU which labels cells in S phase. Arrows indicate ERG1+/BrdU+ cells. (R) Quantification of endothelial cell proliferation (Labeling Index) in untreated (n=4) and atRA control (n=4) and mutants (n=4). Asterisk indicates statistically significant difference (p<0.05) from non-atRA treated control. Scale bars = 500 μm (C-F), 50 μm (G-J), 100μm (N-Q).

Article Snippet: Pregnant dams carrying control and Foxc1 hith/lacz embryos were fed an all trans-retinoic acid (atRA) enriched diet (200 mg/Kg of food; Harlan Teklab Custom diets) from E10 to E14.5 and sacrificed on E14.5 for harvesting control and mutant tissue.

Techniques: Control, Mutagenesis, Immunohistochemistry, Derivative Assay, Labeling, Marker

Journal: Developmental biology

Article Title: Cerebrovascular defects in Foxc1 mutants correlate with aberrant WNT and VEGF-A pathways downstream of retinoic acid from the meninges

doi: 10.1016/j.ydbio.2016.09.019

Figure Lengend Snippet: (A-D) Immunolabeling for β-galactosidase (β-gal) (green) and IB4 (red) of E14.5 coronal brain sections showing PNVP of the cerebral cortex from Foxc1+/+ and Foxc1hith/lacZ (Foxc1h/l) carrying the WNT reporter transgene, Bat-gal-LacZ. (E) Quantification of Bat-gal+ endothelial cells in PNVP per 20x field (n=3). Arrows indicate Bat-gal+/IB4+ endothelial cells. Asterisks indicate statistically significant decrease (p<0.05) from non-atRA control and hashtag (#) indicates a statistically significant increase (p<0.05) from a non-atRA treated control. (F) Approximate timeline PVVP and PNVP growth in the neocortex and timing of tamoxifen (tmx) injections for Pdgfbi-Cre;Ctnnb1-flox experiment. (G,H) E14.5 neocortex labeled with GLUT-1 from control (Ctnnb1-fl/fl) and mutant (Pdgfbi-Cre;Ctnnb1-fl/fl). Arrows indicate PNVP-derived vessels entering cerebral wall. (I) Quantification of number of blood vessels entering cerebral cortex from PNVP. Asterisks indicate statistically significant difference (p<0.05; n=3) from control. (J) Graph depicts gene expression of WNT inhibitors in E14.5 telencephalic meninges/PNVP from atRA-treated control (n=3) and mutants (Foxc1hith/lacZ) (n=3) relative to non-atRA treat control (n=3). Asterisks indicate statistically significant difference (p<0.05) from non-RA treated control. Hashtag (#) indicates statistically significant difference (p<0.05) from non-atRA Foxc1hith/lacZ mutant sample. Scale bars = 100 μm.

Article Snippet: Pregnant dams carrying control and Foxc1 hith/lacz embryos were fed an all trans-retinoic acid (atRA) enriched diet (200 mg/Kg of food; Harlan Teklab Custom diets) from E10 to E14.5 and sacrificed on E14.5 for harvesting control and mutant tissue.

Techniques: Immunolabeling, Control, Labeling, Mutagenesis, Derivative Assay, Gene Expression

Journal: Developmental biology

Article Title: Cerebrovascular defects in Foxc1 mutants correlate with aberrant WNT and VEGF-A pathways downstream of retinoic acid from the meninges

doi: 10.1016/j.ydbio.2016.09.019

Figure Lengend Snippet: (A-D) β-galactosidase (β-gal) (green) and IB4 (red) labeling of E14.5 coronal brain sections depicting the intracortical vasculature from Foxc1+/+ and Foxc1hith/lacZ (Foxc1h/l) carrying the WNT reporter transgene, Bat-gal-LacZ. (E-J) In situs of Wnt7a and Wnt7b on E14.5 wildtype and Foxc1hith/lacZ mutant untreated or exposed to atRA from E10-E14.5. CP: cortical plate, IZ: intermediate zone, SZ: subventricular zone, VZ: ventricular zone. (K) WNT7A and WNT7B protein levels in E14.5 cortical lysates from non-untreated (n=3) and atRA treated control (n=3) and mutants (Foxc1hith/lacZ) (n=3) quantified via ELISA. (L, M) WNT7A and WNT7B protein expression in lysates from non-treated (n=3), atRA-treated (n=3) and atRA+pan-RAR (n=3) antagonist treated cultured cortical progenitors (L) and neurons (N). Asterisks indicate statistically significant difference (p<0.05) from non-RA treated control cells. Hashtag (#) indicates statistically significant difference (p<0.05) from atRA treated cells. Scale bars = 100 μm.

Article Snippet: Pregnant dams carrying control and Foxc1 hith/lacz embryos were fed an all trans-retinoic acid (atRA) enriched diet (200 mg/Kg of food; Harlan Teklab Custom diets) from E10 to E14.5 and sacrificed on E14.5 for harvesting control and mutant tissue.

Techniques: Labeling, Mutagenesis, Control, Enzyme-linked Immunosorbent Assay, Expressing, Cell Culture

Journal: Developmental biology

Article Title: Cerebrovascular defects in Foxc1 mutants correlate with aberrant WNT and VEGF-A pathways downstream of retinoic acid from the meninges

doi: 10.1016/j.ydbio.2016.09.019

Figure Lengend Snippet: (A) VEGF-A and (B) VEGF-C protein levels in E14.5 cortical lysates from non-treated (n=3) and atRA treated control (n=4) and mutants (Foxc1hith/lacZ) (n=3) quantified via ELISA. Asterisks indicate statistically significant difference (p<0.05) from non-RA treated control (n=4). Hashtag (#) indicates statistically significant difference (p<0.05) from both non-RA treated control and Foxc1hith/lacZ mutant samples. (C) Vegfa gene expression in hypoxia (n=3), atRA (n=3) or atRA+pan-RAR antagonist (n=3) treated E14 cortical progenitor cells relative to Vegfa expression of control (untreated E14 cortical progenitor cells) (n=3) measured by real time-PCR. (D) VEGF-A protein levels in conditioned medium collected from control (untreated) (n=3), atRA treated (n=3) and RA+pan-RAR antagonist (n=3) treated mouse E14 cortical progenitor cells.

Article Snippet: Pregnant dams carrying control and Foxc1 hith/lacz embryos were fed an all trans-retinoic acid (atRA) enriched diet (200 mg/Kg of food; Harlan Teklab Custom diets) from E10 to E14.5 and sacrificed on E14.5 for harvesting control and mutant tissue.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Mutagenesis, Gene Expression, Expressing, Real-time Polymerase Chain Reaction