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bvos  (Cambridge Isotope Laboratories)
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Cambridge Isotope Laboratories bvos
a CRISPR-Cas9 genome editing was used to generate PFKFB3 knockout iPS cells. Immunofluorescence confocal imaging of unedited control (CTR) and PFKFB3 knockout (P206, P208) BVO sections showing pericyte coverage (PDGFRβ, magenta) of CD31 + ECs (green). b Percentage of pericyte coverage CTR n = 6 <t>BVOs,</t> P206 n = 7 BVOs, P208 n = 7 BVOs, from three separate preparations. One-two sections per BVO were assessed. c Quantification of vessel density and ( d ) length in CTR n = 6 BVOs, P206 n = 7 BVOs, P208 n = 7 BVOs, from three separate preparations in four different areas per x10 images. One-two sections per BVO were assessed. Values are presented as mean ± SD using two-way ANOVA followed by Tukey’s multiple comparisons tests. ( b : CTR vs. P206 **** p = 0.000000000522; CTR vs. P208 **** p = 0.000000128; c : * p = 0.0374; CTR vs P206 **** p = 0.000027; CTR vs P208 **** p = 0.000000022; d : CTR vs P206 **** p = 0.000088; CTR vs P208 **** p = 0.000005). ns not significant. Bar scales 200 and 50 μm. Effect of PFKFB3 knockout on 13 C label incorporation into metabolites related to ( e ) glycolysis, ( f ) the TCA cycle and ( g ) glycolytic branch pathways after 3 h <t>of</t> <t>incubation</t> with 13 C 6 -glucose. Data represents mean ± SEM, n = 4 independent experiments, statistical significance was assessed by a two-way ANOVA with Holm-Sidak post-hoc test. ( e : LAC ** p = 0.00618; PEP ** p = 0.00531; DHAP * p = 0.02990; G3P **** p = 0.000000798; FBP * p = 0.02749; F6P * p = 0.04479; G6P* p = 0.00473). G6P glucose 6-phosphate, F6P fructose 6-phosphate, FBP fructose 1,6-bisphosphate, G3P glyceraldehyde 3-phosphate, DHAP dihydroxyacetone phosphate, 1,3-BPG 1,3-bisphosphoglycerate, 2-PG 2-phosphoglycerate, PEP phosphoenolpyruvate, LAC lactate, UDP uridine diphosphate, GlcNAc N -acetylglucosamine, Ru5P ribulose 5-phosphate, R5P ribose 5-phosphate, S7P sedoheptulose-7-phosphate, m + x mass isotopologues x. Source data are provided as a Source Data file.
Bvos, supplied by Cambridge Isotope Laboratories, 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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a CRISPR-Cas9 genome editing was used to generate PFKFB3 knockout iPS cells. Immunofluorescence confocal imaging of unedited control (CTR) and PFKFB3 knockout (P206, P208) BVO sections showing pericyte coverage (PDGFRβ, magenta) of CD31 + ECs (green). b Percentage of pericyte coverage CTR n = 6 BVOs, P206 n = 7 BVOs, P208 n = 7 BVOs, from three separate preparations. One-two sections per BVO were assessed. c Quantification of vessel density and ( d ) length in CTR n = 6 BVOs, P206 n = 7 BVOs, P208 n = 7 BVOs, from three separate preparations in four different areas per x10 images. One-two sections per BVO were assessed. Values are presented as mean ± SD using two-way ANOVA followed by Tukey’s multiple comparisons tests. ( b : CTR vs. P206 **** p = 0.000000000522; CTR vs. P208 **** p = 0.000000128; c : * p = 0.0374; CTR vs P206 **** p = 0.000027; CTR vs P208 **** p = 0.000000022; d : CTR vs P206 **** p = 0.000088; CTR vs P208 **** p = 0.000005). ns not significant. Bar scales 200 and 50 μm. Effect of PFKFB3 knockout on 13 C label incorporation into metabolites related to ( e ) glycolysis, ( f ) the TCA cycle and ( g ) glycolytic branch pathways after 3 h of incubation with 13 C 6 -glucose. Data represents mean ± SEM, n = 4 independent experiments, statistical significance was assessed by a two-way ANOVA with Holm-Sidak post-hoc test. ( e : LAC ** p = 0.00618; PEP ** p = 0.00531; DHAP * p = 0.02990; G3P **** p = 0.000000798; FBP * p = 0.02749; F6P * p = 0.04479; G6P* p = 0.00473). G6P glucose 6-phosphate, F6P fructose 6-phosphate, FBP fructose 1,6-bisphosphate, G3P glyceraldehyde 3-phosphate, DHAP dihydroxyacetone phosphate, 1,3-BPG 1,3-bisphosphoglycerate, 2-PG 2-phosphoglycerate, PEP phosphoenolpyruvate, LAC lactate, UDP uridine diphosphate, GlcNAc N -acetylglucosamine, Ru5P ribulose 5-phosphate, R5P ribose 5-phosphate, S7P sedoheptulose-7-phosphate, m + x mass isotopologues x. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Human blood vessel organoids reveal a critical role for CTGF in maintaining microvascular integrity

doi: 10.1038/s41467-023-41326-2

Figure Lengend Snippet: a CRISPR-Cas9 genome editing was used to generate PFKFB3 knockout iPS cells. Immunofluorescence confocal imaging of unedited control (CTR) and PFKFB3 knockout (P206, P208) BVO sections showing pericyte coverage (PDGFRβ, magenta) of CD31 + ECs (green). b Percentage of pericyte coverage CTR n = 6 BVOs, P206 n = 7 BVOs, P208 n = 7 BVOs, from three separate preparations. One-two sections per BVO were assessed. c Quantification of vessel density and ( d ) length in CTR n = 6 BVOs, P206 n = 7 BVOs, P208 n = 7 BVOs, from three separate preparations in four different areas per x10 images. One-two sections per BVO were assessed. Values are presented as mean ± SD using two-way ANOVA followed by Tukey’s multiple comparisons tests. ( b : CTR vs. P206 **** p = 0.000000000522; CTR vs. P208 **** p = 0.000000128; c : * p = 0.0374; CTR vs P206 **** p = 0.000027; CTR vs P208 **** p = 0.000000022; d : CTR vs P206 **** p = 0.000088; CTR vs P208 **** p = 0.000005). ns not significant. Bar scales 200 and 50 μm. Effect of PFKFB3 knockout on 13 C label incorporation into metabolites related to ( e ) glycolysis, ( f ) the TCA cycle and ( g ) glycolytic branch pathways after 3 h of incubation with 13 C 6 -glucose. Data represents mean ± SEM, n = 4 independent experiments, statistical significance was assessed by a two-way ANOVA with Holm-Sidak post-hoc test. ( e : LAC ** p = 0.00618; PEP ** p = 0.00531; DHAP * p = 0.02990; G3P **** p = 0.000000798; FBP * p = 0.02749; F6P * p = 0.04479; G6P* p = 0.00473). G6P glucose 6-phosphate, F6P fructose 6-phosphate, FBP fructose 1,6-bisphosphate, G3P glyceraldehyde 3-phosphate, DHAP dihydroxyacetone phosphate, 1,3-BPG 1,3-bisphosphoglycerate, 2-PG 2-phosphoglycerate, PEP phosphoenolpyruvate, LAC lactate, UDP uridine diphosphate, GlcNAc N -acetylglucosamine, Ru5P ribulose 5-phosphate, R5P ribose 5-phosphate, S7P sedoheptulose-7-phosphate, m + x mass isotopologues x. Source data are provided as a Source Data file.

Article Snippet: For BVOs, a 3 h incubation with 5 mM U- 13 C 6 -glucose (Cambridge Isotope Laboratories) in DMEM without glucose and pyruvate (Thermo Fisher Scientific) supplemented with 1% penicillin-streptomycin and 1% l -glutamine was performed.

Techniques: CRISPR, Knock-Out, Immunofluorescence, Imaging, Control, Incubation