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  • 99
    Millipore skin cryosections
    Skin Cryosections, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Leica Biosystems cryosection
    Cryosection, supplied by Leica Biosystems, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Leica Microsystems cryosection
    Cadherin 7-expressing BC cells are located at both the lateral and ventral exit points in chick embryos. Cervical spinal cord-containing transverse <t>cryosections</t> derived from E4.5 chick embryos were doubled-labeled with either anti-Ben and anti-Laminin ( Left ) or anti-Cadherin 7 and anti-NF ( Middle , Right ), and the appropriate secondary antibodies. ( Left ) BEN is expressed by both SACMN and vMN at this developmental stage, the FP and the SAN, which is positioned outside and adjacent to the spinal cord. Anti-Laminin labeling demarcates the margin of the spinal cord. ( Middle ) Cadherin 7-expressing BC cells are located at both the LEP and VEP. In this panel, the SAN is labeled by anti-NF. ( Right ) A higher magnification view of the boxed area in the middle panel. SACMN, spinal accessory motor neurons; SAN, spinal accessory nerve; vMN, ventral motor neurons; FP, floor plate; LEP, lateral exit point; VEP, ventral exit point; BC cells, boundary cap cells. Scale bar in middle panel, 100μm, applies to the left and middle panels. Scale bar in right panel, 50 μm.
    Cryosection, supplied by Leica Microsystems, used in various techniques. Bioz Stars score: 91/100, based on 204 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher cryosections
    Validation of a newly generated MCT14 antibody. Renal homogenates were incubated with antibodies against MCT14 (A) and NaPi-IIa (B) in the presence and absence of the antigenic peptide as indicated. Lysates of Xenopus laevis oocytes injected with MCT14 cRNA and non-injected oocytes (n.i.) were incubated with the MCT14 antibody (C); renal homogenate was loaded in the last lane (+). Deglycosylated (+ PNGase) and native (- PNGase) renal homogenates were incubated with antibodies against MCT14 and NaPi-IIa (D). Immunofluorescence was performed on kidney <t>cryosections</t> in the presence and absence of the antigenic peptide (E). Immunofluorescence in oocytes expressing MCT14 and corresponding non-injected oocytes (F).
    Cryosections, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 5494 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Carl Zeiss cryosections
    Adult Myogenin mutants have reduced muscle with more but smaller myofibres. a Myog mutant and sib at 120 dpf from myog kg128/+ incross. Bar = 1 cm. Representative images n = 5 mutants, n = 23 sibs. b Myog kg128 or myog kg125 but not myog f fh265 showed reduced standard weight compared to co-reared sibs at 120 dpf. Dots represent individuals. c Laminin immunodetection on <t>cryosections</t> from 120 dpf myog 128/+ incross. Bar = 100 µm. Representative images, n = 3. d – f Number of muscle fibres in 0.1 mm 2 of adult muscle is increased in mutants ( d ), whereas myofibre cross-sectional area (CSA) is decreased ( e ) reflecting a shift in CSA frequency distribution compared to sibs. g Fewer myonuclear profiles were present within laminin profiles in adult muscle cross-sections in mutants than in sibs, measured from 107 to 490 fibres at similar medio-lateral and dorso-ventral positions of trunk muscle of three fish per genotype. Mean ± SEM, t test. h Proportions of muscle fibres with indicated number of myonuclei within fibre cross-sectional profile. In sibs, > 90% of fibres have more than one nuclear profile, compared with
    Cryosections, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 94/100, based on 1596 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Leica Microsystems cryosectioning
    Adult Myogenin mutants have reduced muscle with more but smaller myofibres. a Myog mutant and sib at 120 dpf from myog kg128/+ incross. Bar = 1 cm. Representative images n = 5 mutants, n = 23 sibs. b Myog kg128 or myog kg125 but not myog f fh265 showed reduced standard weight compared to co-reared sibs at 120 dpf. Dots represent individuals. c Laminin immunodetection on <t>cryosections</t> from 120 dpf myog 128/+ incross. Bar = 100 µm. Representative images, n = 3. d – f Number of muscle fibres in 0.1 mm 2 of adult muscle is increased in mutants ( d ), whereas myofibre cross-sectional area (CSA) is decreased ( e ) reflecting a shift in CSA frequency distribution compared to sibs. g Fewer myonuclear profiles were present within laminin profiles in adult muscle cross-sections in mutants than in sibs, measured from 107 to 490 fibres at similar medio-lateral and dorso-ventral positions of trunk muscle of three fish per genotype. Mean ± SEM, t test. h Proportions of muscle fibres with indicated number of myonuclei within fibre cross-sectional profile. In sibs, > 90% of fibres have more than one nuclear profile, compared with
    Cryosectioning, supplied by Leica Microsystems, used in various techniques. Bioz Stars score: 90/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher cryosectioning
    Adult Myogenin mutants have reduced muscle with more but smaller myofibres. a Myog mutant and sib at 120 dpf from myog kg128/+ incross. Bar = 1 cm. Representative images n = 5 mutants, n = 23 sibs. b Myog kg128 or myog kg125 but not myog f fh265 showed reduced standard weight compared to co-reared sibs at 120 dpf. Dots represent individuals. c Laminin immunodetection on <t>cryosections</t> from 120 dpf myog 128/+ incross. Bar = 100 µm. Representative images, n = 3. d – f Number of muscle fibres in 0.1 mm 2 of adult muscle is increased in mutants ( d ), whereas myofibre cross-sectional area (CSA) is decreased ( e ) reflecting a shift in CSA frequency distribution compared to sibs. g Fewer myonuclear profiles were present within laminin profiles in adult muscle cross-sections in mutants than in sibs, measured from 107 to 490 fibres at similar medio-lateral and dorso-ventral positions of trunk muscle of three fish per genotype. Mean ± SEM, t test. h Proportions of muscle fibres with indicated number of myonuclei within fibre cross-sectional profile. In sibs, > 90% of fibres have more than one nuclear profile, compared with
    Cryosectioning, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Leica Microsystems cryosection machine
    Adult Myogenin mutants have reduced muscle with more but smaller myofibres. a Myog mutant and sib at 120 dpf from myog kg128/+ incross. Bar = 1 cm. Representative images n = 5 mutants, n = 23 sibs. b Myog kg128 or myog kg125 but not myog f fh265 showed reduced standard weight compared to co-reared sibs at 120 dpf. Dots represent individuals. c Laminin immunodetection on <t>cryosections</t> from 120 dpf myog 128/+ incross. Bar = 100 µm. Representative images, n = 3. d – f Number of muscle fibres in 0.1 mm 2 of adult muscle is increased in mutants ( d ), whereas myofibre cross-sectional area (CSA) is decreased ( e ) reflecting a shift in CSA frequency distribution compared to sibs. g Fewer myonuclear profiles were present within laminin profiles in adult muscle cross-sections in mutants than in sibs, measured from 107 to 490 fibres at similar medio-lateral and dorso-ventral positions of trunk muscle of three fish per genotype. Mean ± SEM, t test. h Proportions of muscle fibres with indicated number of myonuclei within fibre cross-sectional profile. In sibs, > 90% of fibres have more than one nuclear profile, compared with
    Cryosection Machine, supplied by Leica Microsystems, 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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    Leica Biosystems cryosectioning system
    Adult Myogenin mutants have reduced muscle with more but smaller myofibres. a Myog mutant and sib at 120 dpf from myog kg128/+ incross. Bar = 1 cm. Representative images n = 5 mutants, n = 23 sibs. b Myog kg128 or myog kg125 but not myog f fh265 showed reduced standard weight compared to co-reared sibs at 120 dpf. Dots represent individuals. c Laminin immunodetection on <t>cryosections</t> from 120 dpf myog 128/+ incross. Bar = 100 µm. Representative images, n = 3. d – f Number of muscle fibres in 0.1 mm 2 of adult muscle is increased in mutants ( d ), whereas myofibre cross-sectional area (CSA) is decreased ( e ) reflecting a shift in CSA frequency distribution compared to sibs. g Fewer myonuclear profiles were present within laminin profiles in adult muscle cross-sections in mutants than in sibs, measured from 107 to 490 fibres at similar medio-lateral and dorso-ventral positions of trunk muscle of three fish per genotype. Mean ± SEM, t test. h Proportions of muscle fibres with indicated number of myonuclei within fibre cross-sectional profile. In sibs, > 90% of fibres have more than one nuclear profile, compared with
    Cryosectioning System, supplied by Leica Biosystems, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    OriGene murine melanoma cryosections human melanoma cryosections
    CBD-CCL4 exhibits high affinity to collagen and accumulates in tumor following intravenous injection. ( A ) WT CCL4 and CBD-CCL4 were analyzed by SDS-PAGE followed by Coomassie blue staining. ( B and C ) Affinity of CBD-CCL4 against (B) collagen I and (C) collagen III was measured by SPR. SPR chips were functionalized with collagen I [~500 resonance units (RU)] and collagen III (~700 RU), and CBD-CCL4 was flowed over the chips at indicated concentrations. Curves represent the obtained specific responses (in resonance units) to CBD-CCL4. Experimental curves were fitted with 1:1 Langmuir fit model. Binding kinetics values [dissociation constants ( K d ) and rate constants ( k on and k off )] determined from the fitted curves are shown. ( D and E ) Binding of (D) WT CCL4 or (E) CBD-CCL4 to human melanoma <t>cryosections</t> as determined by immunofluorescence microscopy. Scale bars, 100 μM. DAPI, 4′,6-diamidino-2-phenylindole. ( F ) GPCR activation assay comparing signaling of WT CCL4 and CBD-CCL4 in THP1 monocytes. Median effective concentration (EC 50 ) values were calculated using a nonlinear dose-response curve fit model. Each point represents mean ± SEM, n = 3. ( G ) Blood plasma pharmacokinetics was analyzed using DyLight 800–labeled WT CCL4 or CBD-CCL4 in B16F10 melanoma. Four days after tumor inoculation, mice were administered 25 μg of WT CCL4 or the molar equivalent of CBD-CCL4 (25 μg of CCL4 basis or 93 μg of CBD-CCL4) via intravenous injection. Blood was collected at the indicated time points, and plasma was separated and analyzed for CCL4 concentration. Each point represents mean ± SEM, n = 4. ( H ) Biodistribution was analyzed using DyLight 647–labeled WT CCL4 or CBD-CCL4 in EMT6 breast cancer. When the tumor volume reached 500 mm 3 , 25 μg of WT CCL4 or the molar equivalent of CBD-CCL4 (25 μg of CCL4 basis or 93 μg of CBD-CCL4) was given via intravenous injection. Fluorescence intensity in each tumor was measured using an in vivo imaging system (IVIS), converted to percent injected dose using a known standard series, and normalized to the weight of the tumor. Each bar represents mean ± SEM, n = 3. ** P
    Murine Melanoma Cryosections Human Melanoma Cryosections, supplied by OriGene, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam tumor cryosections
    Endocytosis of EGF-Rh in flank and tongue tumors. Mice harboring flank ( A ) or tongue ( B ) HSC3/EGFR-GFP xenografts were injected with EGF-Rh as in Figure 6 . Tumors were dissected 1 hr after injection and fixed in paraformaldehyde. Confocal imaging of <t>cryosections</t> was performed through 405 nm (Hoescht; blue ), 488 nm (EGFR-GFP; green ) and 561 nm (EGF-Rh; red ) channels. Representative montage images of the large areas of tumors are presented. Regions corresponding to images presented in Figure 6 are marked by white rectangles. Scale bars, 25 µm.
    Tumor Cryosections, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher cryosection medium
    Endocytosis of EGF-Rh in flank and tongue tumors. Mice harboring flank ( A ) or tongue ( B ) HSC3/EGFR-GFP xenografts were injected with EGF-Rh as in Figure 6 . Tumors were dissected 1 hr after injection and fixed in paraformaldehyde. Confocal imaging of <t>cryosections</t> was performed through 405 nm (Hoescht; blue ), 488 nm (EGFR-GFP; green ) and 561 nm (EGF-Rh; red ) channels. Representative montage images of the large areas of tumors are presented. Regions corresponding to images presented in Figure 6 are marked by white rectangles. Scale bars, 25 µm.
    Cryosection Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Roche cryosection staining
    Endocytosis of EGF-Rh in flank and tongue tumors. Mice harboring flank ( A ) or tongue ( B ) HSC3/EGFR-GFP xenografts were injected with EGF-Rh as in Figure 6 . Tumors were dissected 1 hr after injection and fixed in paraformaldehyde. Confocal imaging of <t>cryosections</t> was performed through 405 nm (Hoescht; blue ), 488 nm (EGFR-GFP; green ) and 561 nm (EGF-Rh; red ) channels. Representative montage images of the large areas of tumors are presented. Regions corresponding to images presented in Figure 6 are marked by white rectangles. Scale bars, 25 µm.
    Cryosection Staining, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Agilent technologies cryosection
    Blocking VEGF-A using tinzaparin reduces VWF fiber formation in tumor microvasculature. Immunofluorescence stainings for VWF (green) and anti-CD31 (red) in <t>cryosections</t> of ret transgenic tumors were performed. Nuclei were stained with DAPI. Tumor microvessels of control mice showed formation of ULVWF fibers in the vessel lumen (A, arrows). By contrast, microvessels of tinzaparin-treated mice showed almost no ULVWF fiber formation and a punctual pattern of VWF within the vessel wall (B, arrowheads), indicative of reduced endothelial cell activation. Representative pictures of tumor microvessels are shown (n = 10 animals of 2 independent experiments; scale bars = 20 µm). Tinzaparin treatment correlated with a significant reduction of vessels with intraluminal VWF fibers (C). Tumor vessels were analyzed for platelet aggregation using VWF (green) and GPIb (red) staining. Quantification showed a significant increase in platelet-covered area in the lumen of tumor vessels compared with control. This effect was abolished by treatment with tinzaparin (D-E). Panel Ei shows a single platelet in the lumen of a tumor blood vessel. In addition, the treatment of ret transgenic mice with tinzaparin (gray) reduced the appearance of middle (ii) and big aggregates (iii) to healthy control skin levels (white) compared with vehicle-treated control tumors (black). Plot shows mean ± SD (n = 5-10; * P
    Cryosection, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam cryosections
    CCN1 inhibits hepatic fibrosis by inducing myofibroblast senescence. (A) Adjacent serial liver <t>cryosections</t> from Ccn1 flox / flox and Ccn1 Δ Hep mice treated with CCl 4 to induce fibrosis were stained for SA-β-Gal (upper panels, blue; n = 7)
    Cryosections, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 1803 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ANATECH Corp cryosections
    CCN1 inhibits hepatic fibrosis by inducing myofibroblast senescence. (A) Adjacent serial liver <t>cryosections</t> from Ccn1 flox / flox and Ccn1 Δ Hep mice treated with CCl 4 to induce fibrosis were stained for SA-β-Gal (upper panels, blue; n = 7)
    Cryosections, supplied by ANATECH Corp, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Avantor cryosections
    Sequential Expression of Germ-Cell-Related Transcription Factors in Single Cells during hPGCLC Specification (A and B) Immunofluorscence analysis for (A) BLIMP1, SOX17, and TFAP2C and (B) BLIMP1 and T in <t>cryosections</t> of day 1–8 embryoids after hPGCLC induction. Bottom row in (B) shows high exposure (digital) image of T, indicating low but specific expression in hPGCLC. SOX17-positive or BLIMP1-positive cells are highlighted. Scale bars, 50 μm. (C) Percentage of SOX17-positive (+) cells in day 1–8 embryoids that were also TFAP2C+ or BLIMP1+. Corresponds to data in Figure 4 A. (D) Percentage of BLIMP1-positive (+) cells in day 1–8 embryoids that were TFAP2C+, NANOG+, or OCT4+. Corresponds to data in Figures 4 A, S4 A, and S4B. (E) Summary model for dynamics of hPGCLC specification in embryoids. SOX17-positive cells are first scattered in day 1 embryoids. They gain expression of BLIMP1, TFAP2C, and NANOG sequentially and form a cluster from day 2 onward until the formation of nascent hPGCLC.
    Cryosections, supplied by Avantor, used in various techniques. Bioz Stars score: 92/100, based on 379 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Becton Dickinson cryosections
    MAPC-engrafted mouse with chronic GVHD. Intravital microscopy and histopathological findings in one NOD-SCID mouse that received 0.6 × 10 6 MAPCs 16 wk earlier are consistent with the development of chronic GVHD. Left panels show the presence of GFP + cells in the GVHD target organs, specifically skin (underlayer), liver, lung, and ileum. All intravital images were taken at a zoom factor of 8× and a transfer lens of 0.63× with an MZFLIII stereomicroscope (300 millisecond exposure). Right panels show the corresponding hematoxylin and eosin stains of <t>cryosections</t> taken from the same mouse (bar, 900 μm). The skin shows inflammation in the dermis and subdermis (black arrow), extensive epidermal hyperplasia (white arrow), and dyskeratosis (asterisk). GVHD score, 3.5 (0–4 scale). The liver has moderate inflammation around the portal triads with evidence of bile duct degeneration (black arrow). GVHD score, 3.0. The lung has peribronchiolar inflammation (b, bronchiole) and extensive parenchymal inflammation in surrounding areas with organizing alveolitis and fibrosis. GVHD score, 4.0. The ileum shows only mild inflammation of the lamina propria as is typical for chronic GVHD. GVHD score, 0.5.
    Cryosections, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 1320 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biogenex cryosections
    MAPC-engrafted mouse with chronic GVHD. Intravital microscopy and histopathological findings in one NOD-SCID mouse that received 0.6 × 10 6 MAPCs 16 wk earlier are consistent with the development of chronic GVHD. Left panels show the presence of GFP + cells in the GVHD target organs, specifically skin (underlayer), liver, lung, and ileum. All intravital images were taken at a zoom factor of 8× and a transfer lens of 0.63× with an MZFLIII stereomicroscope (300 millisecond exposure). Right panels show the corresponding hematoxylin and eosin stains of <t>cryosections</t> taken from the same mouse (bar, 900 μm). The skin shows inflammation in the dermis and subdermis (black arrow), extensive epidermal hyperplasia (white arrow), and dyskeratosis (asterisk). GVHD score, 3.5 (0–4 scale). The liver has moderate inflammation around the portal triads with evidence of bile duct degeneration (black arrow). GVHD score, 3.0. The lung has peribronchiolar inflammation (b, bronchiole) and extensive parenchymal inflammation in surrounding areas with organizing alveolitis and fibrosis. GVHD score, 4.0. The ileum shows only mild inflammation of the lamina propria as is typical for chronic GVHD. GVHD score, 0.5.
    Cryosections, supplied by Biogenex, used in various techniques. Bioz Stars score: 93/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc cryosections
    Venules are the source of hemorrhage. A–D: <t>Cryosections</t> immunolabeled for laminin (brown) , which identifies both arterioles and venules, and stained for alkaline phosphatase (blue) , which labels only arterioles, showing hemorrhages in the cortex
    Cryosections, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 471 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Covance cryosections
    Progressive thymic epithelium phenotypes in Foxn1 lacZ mutants . (A-D) <t>Cryosections</t> were stained for Keratin 8 (green) and Keratin 5 (red). Scale bar: 200 μm. Arrowhead, perivascular space; *, cystic structure. (A) Two-week + /lacZ , + /nu , and lacZ/lacZ
    Cryosections, supplied by Covance, used in various techniques. Bioz Stars score: 93/100, based on 152 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Developmental Studies Hybridoma Bank cryosections
    Oral treatment with Ac 4 ManNAc prevents appearance of rimmed vacuoles. A , histochemistry of gastrocnemius <t>cryosections</t> from DMRV/hIBM mice in treated and placebo groups is shown. In the placebo group, H E staining shows variations in fiber size,
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    Image Search Results


    Cadherin 7-expressing BC cells are located at both the lateral and ventral exit points in chick embryos. Cervical spinal cord-containing transverse cryosections derived from E4.5 chick embryos were doubled-labeled with either anti-Ben and anti-Laminin ( Left ) or anti-Cadherin 7 and anti-NF ( Middle , Right ), and the appropriate secondary antibodies. ( Left ) BEN is expressed by both SACMN and vMN at this developmental stage, the FP and the SAN, which is positioned outside and adjacent to the spinal cord. Anti-Laminin labeling demarcates the margin of the spinal cord. ( Middle ) Cadherin 7-expressing BC cells are located at both the LEP and VEP. In this panel, the SAN is labeled by anti-NF. ( Right ) A higher magnification view of the boxed area in the middle panel. SACMN, spinal accessory motor neurons; SAN, spinal accessory nerve; vMN, ventral motor neurons; FP, floor plate; LEP, lateral exit point; VEP, ventral exit point; BC cells, boundary cap cells. Scale bar in middle panel, 100μm, applies to the left and middle panels. Scale bar in right panel, 50 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Crossing the Border: Molecular Control of Motor Axon Exit

    doi: 10.3390/ijms12128539

    Figure Lengend Snippet: Cadherin 7-expressing BC cells are located at both the lateral and ventral exit points in chick embryos. Cervical spinal cord-containing transverse cryosections derived from E4.5 chick embryos were doubled-labeled with either anti-Ben and anti-Laminin ( Left ) or anti-Cadherin 7 and anti-NF ( Middle , Right ), and the appropriate secondary antibodies. ( Left ) BEN is expressed by both SACMN and vMN at this developmental stage, the FP and the SAN, which is positioned outside and adjacent to the spinal cord. Anti-Laminin labeling demarcates the margin of the spinal cord. ( Middle ) Cadherin 7-expressing BC cells are located at both the LEP and VEP. In this panel, the SAN is labeled by anti-NF. ( Right ) A higher magnification view of the boxed area in the middle panel. SACMN, spinal accessory motor neurons; SAN, spinal accessory nerve; vMN, ventral motor neurons; FP, floor plate; LEP, lateral exit point; VEP, ventral exit point; BC cells, boundary cap cells. Scale bar in middle panel, 100μm, applies to the left and middle panels. Scale bar in right panel, 50 μm.

    Article Snippet: Cryosections (16 μm) were generated using a Leica cryostat (model #; CM3050 S; Leica Microsystems) and collected on Superfrost Plus microscope slides (Fisher Scientific, Cat. No. 12-550-15).

    Techniques: Expressing, Derivative Assay, Labeling

    Validation of a newly generated MCT14 antibody. Renal homogenates were incubated with antibodies against MCT14 (A) and NaPi-IIa (B) in the presence and absence of the antigenic peptide as indicated. Lysates of Xenopus laevis oocytes injected with MCT14 cRNA and non-injected oocytes (n.i.) were incubated with the MCT14 antibody (C); renal homogenate was loaded in the last lane (+). Deglycosylated (+ PNGase) and native (- PNGase) renal homogenates were incubated with antibodies against MCT14 and NaPi-IIa (D). Immunofluorescence was performed on kidney cryosections in the presence and absence of the antigenic peptide (E). Immunofluorescence in oocytes expressing MCT14 and corresponding non-injected oocytes (F).

    Journal: PLoS ONE

    Article Title: Renal localization and regulation by dietary phosphate of the MCT14 orphan transporter

    doi: 10.1371/journal.pone.0177942

    Figure Lengend Snippet: Validation of a newly generated MCT14 antibody. Renal homogenates were incubated with antibodies against MCT14 (A) and NaPi-IIa (B) in the presence and absence of the antigenic peptide as indicated. Lysates of Xenopus laevis oocytes injected with MCT14 cRNA and non-injected oocytes (n.i.) were incubated with the MCT14 antibody (C); renal homogenate was loaded in the last lane (+). Deglycosylated (+ PNGase) and native (- PNGase) renal homogenates were incubated with antibodies against MCT14 and NaPi-IIa (D). Immunofluorescence was performed on kidney cryosections in the presence and absence of the antigenic peptide (E). Immunofluorescence in oocytes expressing MCT14 and corresponding non-injected oocytes (F).

    Article Snippet: Cryosections of 5 μm were mounted on slides (Superfrost Plus, Thermo Scientific) and blocked with 3% bovine serum albumin/PBS; the primary antibodies against MCT14 (1/1000), NKCC2 [ ] (1/1000), Uromodulin [ ] (1/400), NCC [ ] (1/500) or NaPi-IIa [ ] (1/400) were diluted in PBS and added to the cryosections for incubation over night at 4°C.

    Techniques: Generated, Incubation, Injection, Immunofluorescence, Expressing

    Confocal images illustrating intestine cryosections 1 week after transplantation. a Duodenal cryosection from a control (nontransplanted) piglet. b Cryosection from the duodenum of a piglet sacrificed 1 week after transplantation of PKH26-labeled WJCs by IP injection. Numerous PKH26-labeled (red) cells are visible in the stromal layer underlying the epithelium (arrows). s submucosal layer, lp lamina propria, vi villus

    Journal: Stem Cell Research & Therapy

    Article Title: Porcine Wharton’s jelly cells distribute throughout the body after intraperitoneal injection

    doi: 10.1186/s13287-018-0775-7

    Figure Lengend Snippet: Confocal images illustrating intestine cryosections 1 week after transplantation. a Duodenal cryosection from a control (nontransplanted) piglet. b Cryosection from the duodenum of a piglet sacrificed 1 week after transplantation of PKH26-labeled WJCs by IP injection. Numerous PKH26-labeled (red) cells are visible in the stromal layer underlying the epithelium (arrows). s submucosal layer, lp lamina propria, vi villus

    Article Snippet: Tissues were preserved in DMSO/EDTA/saturated NaCl buffer (DESS) buffer and 4% paraformaldehyde solution (16%; Thermo Scientific) for cryosection and DNA isolation.

    Techniques: Transplantation Assay, Labeling, Injection

    Confocal images of cecal and colon cryosections from 1-week-old piglets given cells IP at birth. a Cryosection from the cecum of a piglet receiving PKH26-labeled WJCs. b The colon from a control pig and c colon of 1-week-old piglet after IP injection of PKH26-labeled WJCs at birth. Arrows indicate PKH26-labeled (red) cells. lp lamina propria

    Journal: Stem Cell Research & Therapy

    Article Title: Porcine Wharton’s jelly cells distribute throughout the body after intraperitoneal injection

    doi: 10.1186/s13287-018-0775-7

    Figure Lengend Snippet: Confocal images of cecal and colon cryosections from 1-week-old piglets given cells IP at birth. a Cryosection from the cecum of a piglet receiving PKH26-labeled WJCs. b The colon from a control pig and c colon of 1-week-old piglet after IP injection of PKH26-labeled WJCs at birth. Arrows indicate PKH26-labeled (red) cells. lp lamina propria

    Article Snippet: Tissues were preserved in DMSO/EDTA/saturated NaCl buffer (DESS) buffer and 4% paraformaldehyde solution (16%; Thermo Scientific) for cryosection and DNA isolation.

    Techniques: Labeling, Injection

    Adult Myogenin mutants have reduced muscle with more but smaller myofibres. a Myog mutant and sib at 120 dpf from myog kg128/+ incross. Bar = 1 cm. Representative images n = 5 mutants, n = 23 sibs. b Myog kg128 or myog kg125 but not myog f fh265 showed reduced standard weight compared to co-reared sibs at 120 dpf. Dots represent individuals. c Laminin immunodetection on cryosections from 120 dpf myog 128/+ incross. Bar = 100 µm. Representative images, n = 3. d – f Number of muscle fibres in 0.1 mm 2 of adult muscle is increased in mutants ( d ), whereas myofibre cross-sectional area (CSA) is decreased ( e ) reflecting a shift in CSA frequency distribution compared to sibs. g Fewer myonuclear profiles were present within laminin profiles in adult muscle cross-sections in mutants than in sibs, measured from 107 to 490 fibres at similar medio-lateral and dorso-ventral positions of trunk muscle of three fish per genotype. Mean ± SEM, t test. h Proportions of muscle fibres with indicated number of myonuclei within fibre cross-sectional profile. In sibs, > 90% of fibres have more than one nuclear profile, compared with

    Journal: Nature Communications

    Article Title: Myogenin promotes myocyte fusion to balance fibre number and size

    doi: 10.1038/s41467-018-06583-6

    Figure Lengend Snippet: Adult Myogenin mutants have reduced muscle with more but smaller myofibres. a Myog mutant and sib at 120 dpf from myog kg128/+ incross. Bar = 1 cm. Representative images n = 5 mutants, n = 23 sibs. b Myog kg128 or myog kg125 but not myog f fh265 showed reduced standard weight compared to co-reared sibs at 120 dpf. Dots represent individuals. c Laminin immunodetection on cryosections from 120 dpf myog 128/+ incross. Bar = 100 µm. Representative images, n = 3. d – f Number of muscle fibres in 0.1 mm 2 of adult muscle is increased in mutants ( d ), whereas myofibre cross-sectional area (CSA) is decreased ( e ) reflecting a shift in CSA frequency distribution compared to sibs. g Fewer myonuclear profiles were present within laminin profiles in adult muscle cross-sections in mutants than in sibs, measured from 107 to 490 fibres at similar medio-lateral and dorso-ventral positions of trunk muscle of three fish per genotype. Mean ± SEM, t test. h Proportions of muscle fibres with indicated number of myonuclei within fibre cross-sectional profile. In sibs, > 90% of fibres have more than one nuclear profile, compared with

    Article Snippet: Cryosections (15 µm) from three anteroposterior positions were immunolabelled for Laminin and counterstained with Hoechst 33342 as described and three or four images in consistent mediolateral and dorso-ventral somitic areas of sibs and mutants were acquired using an Axiovert 200 M microscope (Zeiss) equipped with LD A-plan ×20/0.85 objective.

    Techniques: Mutagenesis, Immunodetection, Fluorescence In Situ Hybridization

    CBD-CCL4 exhibits high affinity to collagen and accumulates in tumor following intravenous injection. ( A ) WT CCL4 and CBD-CCL4 were analyzed by SDS-PAGE followed by Coomassie blue staining. ( B and C ) Affinity of CBD-CCL4 against (B) collagen I and (C) collagen III was measured by SPR. SPR chips were functionalized with collagen I [~500 resonance units (RU)] and collagen III (~700 RU), and CBD-CCL4 was flowed over the chips at indicated concentrations. Curves represent the obtained specific responses (in resonance units) to CBD-CCL4. Experimental curves were fitted with 1:1 Langmuir fit model. Binding kinetics values [dissociation constants ( K d ) and rate constants ( k on and k off )] determined from the fitted curves are shown. ( D and E ) Binding of (D) WT CCL4 or (E) CBD-CCL4 to human melanoma cryosections as determined by immunofluorescence microscopy. Scale bars, 100 μM. DAPI, 4′,6-diamidino-2-phenylindole. ( F ) GPCR activation assay comparing signaling of WT CCL4 and CBD-CCL4 in THP1 monocytes. Median effective concentration (EC 50 ) values were calculated using a nonlinear dose-response curve fit model. Each point represents mean ± SEM, n = 3. ( G ) Blood plasma pharmacokinetics was analyzed using DyLight 800–labeled WT CCL4 or CBD-CCL4 in B16F10 melanoma. Four days after tumor inoculation, mice were administered 25 μg of WT CCL4 or the molar equivalent of CBD-CCL4 (25 μg of CCL4 basis or 93 μg of CBD-CCL4) via intravenous injection. Blood was collected at the indicated time points, and plasma was separated and analyzed for CCL4 concentration. Each point represents mean ± SEM, n = 4. ( H ) Biodistribution was analyzed using DyLight 647–labeled WT CCL4 or CBD-CCL4 in EMT6 breast cancer. When the tumor volume reached 500 mm 3 , 25 μg of WT CCL4 or the molar equivalent of CBD-CCL4 (25 μg of CCL4 basis or 93 μg of CBD-CCL4) was given via intravenous injection. Fluorescence intensity in each tumor was measured using an in vivo imaging system (IVIS), converted to percent injected dose using a known standard series, and normalized to the weight of the tumor. Each bar represents mean ± SEM, n = 3. ** P

    Journal: Science Advances

    Article Title: Recruitment of CD103+ dendritic cells via tumor-targeted chemokine delivery enhances efficacy of checkpoint inhibitor immunotherapy

    doi: 10.1126/sciadv.aay1357

    Figure Lengend Snippet: CBD-CCL4 exhibits high affinity to collagen and accumulates in tumor following intravenous injection. ( A ) WT CCL4 and CBD-CCL4 were analyzed by SDS-PAGE followed by Coomassie blue staining. ( B and C ) Affinity of CBD-CCL4 against (B) collagen I and (C) collagen III was measured by SPR. SPR chips were functionalized with collagen I [~500 resonance units (RU)] and collagen III (~700 RU), and CBD-CCL4 was flowed over the chips at indicated concentrations. Curves represent the obtained specific responses (in resonance units) to CBD-CCL4. Experimental curves were fitted with 1:1 Langmuir fit model. Binding kinetics values [dissociation constants ( K d ) and rate constants ( k on and k off )] determined from the fitted curves are shown. ( D and E ) Binding of (D) WT CCL4 or (E) CBD-CCL4 to human melanoma cryosections as determined by immunofluorescence microscopy. Scale bars, 100 μM. DAPI, 4′,6-diamidino-2-phenylindole. ( F ) GPCR activation assay comparing signaling of WT CCL4 and CBD-CCL4 in THP1 monocytes. Median effective concentration (EC 50 ) values were calculated using a nonlinear dose-response curve fit model. Each point represents mean ± SEM, n = 3. ( G ) Blood plasma pharmacokinetics was analyzed using DyLight 800–labeled WT CCL4 or CBD-CCL4 in B16F10 melanoma. Four days after tumor inoculation, mice were administered 25 μg of WT CCL4 or the molar equivalent of CBD-CCL4 (25 μg of CCL4 basis or 93 μg of CBD-CCL4) via intravenous injection. Blood was collected at the indicated time points, and plasma was separated and analyzed for CCL4 concentration. Each point represents mean ± SEM, n = 4. ( H ) Biodistribution was analyzed using DyLight 647–labeled WT CCL4 or CBD-CCL4 in EMT6 breast cancer. When the tumor volume reached 500 mm 3 , 25 μg of WT CCL4 or the molar equivalent of CBD-CCL4 (25 μg of CCL4 basis or 93 μg of CBD-CCL4) was given via intravenous injection. Fluorescence intensity in each tumor was measured using an in vivo imaging system (IVIS), converted to percent injected dose using a known standard series, and normalized to the weight of the tumor. Each bar represents mean ± SEM, n = 3. ** P

    Article Snippet: CBD-CCL4 binding to human and murine melanoma cryosections Human melanoma cryosections were obtained from OriGene Technologies.

    Techniques: Injection, SDS Page, Staining, SPR Assay, Binding Assay, Immunofluorescence, Microscopy, Activation Assay, Concentration Assay, Labeling, Mouse Assay, Fluorescence, In Vivo Imaging

    Endocytosis of EGF-Rh in flank and tongue tumors. Mice harboring flank ( A ) or tongue ( B ) HSC3/EGFR-GFP xenografts were injected with EGF-Rh as in Figure 6 . Tumors were dissected 1 hr after injection and fixed in paraformaldehyde. Confocal imaging of cryosections was performed through 405 nm (Hoescht; blue ), 488 nm (EGFR-GFP; green ) and 561 nm (EGF-Rh; red ) channels. Representative montage images of the large areas of tumors are presented. Regions corresponding to images presented in Figure 6 are marked by white rectangles. Scale bars, 25 µm.

    Journal: eLife

    Article Title: EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo

    doi: 10.7554/eLife.31993

    Figure Lengend Snippet: Endocytosis of EGF-Rh in flank and tongue tumors. Mice harboring flank ( A ) or tongue ( B ) HSC3/EGFR-GFP xenografts were injected with EGF-Rh as in Figure 6 . Tumors were dissected 1 hr after injection and fixed in paraformaldehyde. Confocal imaging of cryosections was performed through 405 nm (Hoescht; blue ), 488 nm (EGFR-GFP; green ) and 561 nm (EGF-Rh; red ) channels. Representative montage images of the large areas of tumors are presented. Regions corresponding to images presented in Figure 6 are marked by white rectangles. Scale bars, 25 µm.

    Article Snippet: EEA1 (ab2900) and Y1068 (ab32430) antibodies used for immunostaining of tumor cryosections were from AbCam (Cambridge, MA).

    Techniques: Mouse Assay, Injection, Imaging

    Effects of gefitinib and endocytosis inhibitors on EGF-Rh and EGFR-GFP internalization in tumors. Mice harboring flank HSC3/EGFR-GFP xenografts were administered i.p. with DMSO, gefitinib (30 mg/kg) or Dyngo-4a plus Pitstop2 (1.125 mM/each in 400 μl saline-glucose). 2 hr after these injections, EGF-Rh was i.v. injected as in Figure 6 . Tumors were dissected 1 hr after EGF-Rh injection and fixed in paraformaldehyde. Confocal imaging of cryosections was performed through 405 nm (Hoescht; blue ), 488 nm (EGFR-GFP; green ) and 561 nm (EGF-Rh; red ) channels. Scale bars, 10 µm. Representative merged images are shown. Insets show high-magnification single-channel images of the regions indicated by white rectangles. Intensity scales are identical.

    Journal: eLife

    Article Title: EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo

    doi: 10.7554/eLife.31993

    Figure Lengend Snippet: Effects of gefitinib and endocytosis inhibitors on EGF-Rh and EGFR-GFP internalization in tumors. Mice harboring flank HSC3/EGFR-GFP xenografts were administered i.p. with DMSO, gefitinib (30 mg/kg) or Dyngo-4a plus Pitstop2 (1.125 mM/each in 400 μl saline-glucose). 2 hr after these injections, EGF-Rh was i.v. injected as in Figure 6 . Tumors were dissected 1 hr after EGF-Rh injection and fixed in paraformaldehyde. Confocal imaging of cryosections was performed through 405 nm (Hoescht; blue ), 488 nm (EGFR-GFP; green ) and 561 nm (EGF-Rh; red ) channels. Scale bars, 10 µm. Representative merged images are shown. Insets show high-magnification single-channel images of the regions indicated by white rectangles. Intensity scales are identical.

    Article Snippet: EEA1 (ab2900) and Y1068 (ab32430) antibodies used for immunostaining of tumor cryosections were from AbCam (Cambridge, MA).

    Techniques: Mouse Assay, Injection, Imaging

    Immunofluorescence labeling of EGFR in HSC3/EGFR-GFP tumor flank xenografts. Athymic nude mice harboring HSC3/EGFR-GFP xenograft tumors were dissected, fixed in paraformaldehyde and permeabilized with Triton X-100. Cryosections were labeled with Mab528 followed by secondary Cy3-conjugated antibodies (red). Nuclei were stained with Hoescht. Confocal imaging was performed through 488 nm (GFP, green), 561 nm (Cy3, red) and 405 nm (Hoescht, blue). Scale bar, 10 µm. Insets show high-magnification images of regions indicated by white rectangles. Arrows point on examples of Mab528 and GFP co-localization in clusters and vesicles.

    Journal: eLife

    Article Title: EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo

    doi: 10.7554/eLife.31993

    Figure Lengend Snippet: Immunofluorescence labeling of EGFR in HSC3/EGFR-GFP tumor flank xenografts. Athymic nude mice harboring HSC3/EGFR-GFP xenograft tumors were dissected, fixed in paraformaldehyde and permeabilized with Triton X-100. Cryosections were labeled with Mab528 followed by secondary Cy3-conjugated antibodies (red). Nuclei were stained with Hoescht. Confocal imaging was performed through 488 nm (GFP, green), 561 nm (Cy3, red) and 405 nm (Hoescht, blue). Scale bar, 10 µm. Insets show high-magnification images of regions indicated by white rectangles. Arrows point on examples of Mab528 and GFP co-localization in clusters and vesicles.

    Article Snippet: EEA1 (ab2900) and Y1068 (ab32430) antibodies used for immunostaining of tumor cryosections were from AbCam (Cambridge, MA).

    Techniques: Immunofluorescence, Labeling, Mouse Assay, Staining, Imaging

    Controls for specificity of EEA.1 and pY1068 labeling. Cryosections of HSC3/EGFR-GFP flank tumors were labeled with EEA.1 ( A ) or pY1068 antibodies ( B ), or with non-specific rabbit IgG used in the concentrations equal to the concentrations of specific primary antibodies. Secondary Cy5-conjugated antibodies were used at the same concentration in all samples. Confocal imaging was performed as in Figure 3 . Scale bars, 25 µm.

    Journal: eLife

    Article Title: EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo

    doi: 10.7554/eLife.31993

    Figure Lengend Snippet: Controls for specificity of EEA.1 and pY1068 labeling. Cryosections of HSC3/EGFR-GFP flank tumors were labeled with EEA.1 ( A ) or pY1068 antibodies ( B ), or with non-specific rabbit IgG used in the concentrations equal to the concentrations of specific primary antibodies. Secondary Cy5-conjugated antibodies were used at the same concentration in all samples. Confocal imaging was performed as in Figure 3 . Scale bars, 25 µm.

    Article Snippet: EEA1 (ab2900) and Y1068 (ab32430) antibodies used for immunostaining of tumor cryosections were from AbCam (Cambridge, MA).

    Techniques: Labeling, Concentration Assay, Imaging

    Immunofluorescence labeling of early endosomes and phosphorylated EGFR in HSC3/EGFR-GFP tumor flank xenografts in the presence of EGF-Rh. Athymic nude mice harboring HSC3/EGFR-GFP xenografts were injected with EGF-Rh as in Figure 6 . Tumors were dissected 1 hr after injection and fixed in paraformaldehyde. Cryosections were immunolabelled with EEA1 ( A ) or pY1068 ( B ) antibodies followed by secondary Cy5-conjugated antibodies. Confocal imaging was performed as in Figure 3 . Montage images are shown. Scale bars, 25 µm. Insets show high-magnification images of regions indicated by white rectangles. Arrows point on co-localizations in vesicular compartments.

    Journal: eLife

    Article Title: EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo

    doi: 10.7554/eLife.31993

    Figure Lengend Snippet: Immunofluorescence labeling of early endosomes and phosphorylated EGFR in HSC3/EGFR-GFP tumor flank xenografts in the presence of EGF-Rh. Athymic nude mice harboring HSC3/EGFR-GFP xenografts were injected with EGF-Rh as in Figure 6 . Tumors were dissected 1 hr after injection and fixed in paraformaldehyde. Cryosections were immunolabelled with EEA1 ( A ) or pY1068 ( B ) antibodies followed by secondary Cy5-conjugated antibodies. Confocal imaging was performed as in Figure 3 . Montage images are shown. Scale bars, 25 µm. Insets show high-magnification images of regions indicated by white rectangles. Arrows point on co-localizations in vesicular compartments.

    Article Snippet: EEA1 (ab2900) and Y1068 (ab32430) antibodies used for immunostaining of tumor cryosections were from AbCam (Cambridge, MA).

    Techniques: Immunofluorescence, Labeling, Mouse Assay, Injection, Imaging

    Blocking VEGF-A using tinzaparin reduces VWF fiber formation in tumor microvasculature. Immunofluorescence stainings for VWF (green) and anti-CD31 (red) in cryosections of ret transgenic tumors were performed. Nuclei were stained with DAPI. Tumor microvessels of control mice showed formation of ULVWF fibers in the vessel lumen (A, arrows). By contrast, microvessels of tinzaparin-treated mice showed almost no ULVWF fiber formation and a punctual pattern of VWF within the vessel wall (B, arrowheads), indicative of reduced endothelial cell activation. Representative pictures of tumor microvessels are shown (n = 10 animals of 2 independent experiments; scale bars = 20 µm). Tinzaparin treatment correlated with a significant reduction of vessels with intraluminal VWF fibers (C). Tumor vessels were analyzed for platelet aggregation using VWF (green) and GPIb (red) staining. Quantification showed a significant increase in platelet-covered area in the lumen of tumor vessels compared with control. This effect was abolished by treatment with tinzaparin (D-E). Panel Ei shows a single platelet in the lumen of a tumor blood vessel. In addition, the treatment of ret transgenic mice with tinzaparin (gray) reduced the appearance of middle (ii) and big aggregates (iii) to healthy control skin levels (white) compared with vehicle-treated control tumors (black). Plot shows mean ± SD (n = 5-10; * P

    Journal: Blood

    Article Title: von Willebrand factor fibers promote cancer-associated platelet aggregation in malignant melanoma of mice and humans

    doi: 10.1182/blood-2014-08-595686

    Figure Lengend Snippet: Blocking VEGF-A using tinzaparin reduces VWF fiber formation in tumor microvasculature. Immunofluorescence stainings for VWF (green) and anti-CD31 (red) in cryosections of ret transgenic tumors were performed. Nuclei were stained with DAPI. Tumor microvessels of control mice showed formation of ULVWF fibers in the vessel lumen (A, arrows). By contrast, microvessels of tinzaparin-treated mice showed almost no ULVWF fiber formation and a punctual pattern of VWF within the vessel wall (B, arrowheads), indicative of reduced endothelial cell activation. Representative pictures of tumor microvessels are shown (n = 10 animals of 2 independent experiments; scale bars = 20 µm). Tinzaparin treatment correlated with a significant reduction of vessels with intraluminal VWF fibers (C). Tumor vessels were analyzed for platelet aggregation using VWF (green) and GPIb (red) staining. Quantification showed a significant increase in platelet-covered area in the lumen of tumor vessels compared with control. This effect was abolished by treatment with tinzaparin (D-E). Panel Ei shows a single platelet in the lumen of a tumor blood vessel. In addition, the treatment of ret transgenic mice with tinzaparin (gray) reduced the appearance of middle (ii) and big aggregates (iii) to healthy control skin levels (white) compared with vehicle-treated control tumors (black). Plot shows mean ± SD (n = 5-10; * P

    Article Snippet: Cryosections (10 µm) were incubated with the following primary antibodies: rabbit anti-human VWF (DakoCytomation), rat anti-mouse CD42b (emfret Analytics), mouse anti-human Thrombospondin (Laboratory Vision/Neomarkers), mouse anti-human CD31 (DakoCytomation), rat anti-mouse CD31 (BD Biosciences), Ki67–fluorescein isothiocyanate (FITC) (BD Biosciences), rabbit anti-VEGF-A (Santa Cruz Biotechnology).

    Techniques: Blocking Assay, Immunofluorescence, Transgenic Assay, Staining, Mouse Assay, Activation Assay

    Tinzaparin inhibits VEGF-A–mediated angiogenesis in primary skin tumors and impedes tumor cell metastasis. Tumor-bearing mice were treated with vehicle (A; control) or tinzaparin (B) and cryosections of primary tumors were analyzed by immunofluorescences for CD31. Morphometric quantification of the vessel density (C) demonstrates a significant difference in vessel density upon tinzaparin treatment compared with control tumors. Quantitative assessment of vessels in tinzaparin-treated tumors (D) shows that tinzaparin treatment results in a significant increase of small vessels (

    Journal: Blood

    Article Title: von Willebrand factor fibers promote cancer-associated platelet aggregation in malignant melanoma of mice and humans

    doi: 10.1182/blood-2014-08-595686

    Figure Lengend Snippet: Tinzaparin inhibits VEGF-A–mediated angiogenesis in primary skin tumors and impedes tumor cell metastasis. Tumor-bearing mice were treated with vehicle (A; control) or tinzaparin (B) and cryosections of primary tumors were analyzed by immunofluorescences for CD31. Morphometric quantification of the vessel density (C) demonstrates a significant difference in vessel density upon tinzaparin treatment compared with control tumors. Quantitative assessment of vessels in tinzaparin-treated tumors (D) shows that tinzaparin treatment results in a significant increase of small vessels (

    Article Snippet: Cryosections (10 µm) were incubated with the following primary antibodies: rabbit anti-human VWF (DakoCytomation), rat anti-mouse CD42b (emfret Analytics), mouse anti-human Thrombospondin (Laboratory Vision/Neomarkers), mouse anti-human CD31 (DakoCytomation), rat anti-mouse CD31 (BD Biosciences), Ki67–fluorescein isothiocyanate (FITC) (BD Biosciences), rabbit anti-VEGF-A (Santa Cruz Biotechnology).

    Techniques: Mouse Assay

    Local inhibition of ADAMTS13 promotes VWF fiber formation in microvessels obtained from human melanoma patients. Cryosections of human malignant melanoma tissue, healthy control skin, and basal cell carcinoma were analyzed by immunofluorescence stainings for VWF and CD31 (A-C) or thrombospondin (TSP) (D-E). Nuclei were stained with DAPI. Analysis of healthy skin (A) and human basal cell carcinoma (B) as control demonstrate storage of VWF in the vessel wall (arrowheads). ULVWF fibers are detected in the lumen of the microvessels, correlating to reduced VWF within the vessel wall indicative of EC activation (C, arrows). These ULVWF fibers bind platelets (D, arrows) and are associated with microthrombi formation in distinct microvessels (E, asterisk; n = 5 to 6; scale bars = 20 µm). Quantification showed significantly increased numbers of vessels with luminal VWF fibers in tumor vasculature compared with healthy skin (F). Systemic VWF level in blood samples of malignant melanoma patients was increased compared with healthy control (G). By contrast, only a slight reduction of ADAMTS13 activity was observed (H). Tumor-derived cytokines and growth factors were measured in healthy control skin and tumor of human malignant melanoma by bio-plex. Cytokine levels of IFN-γ (I), TNF-α (J), and IL-6 (K) were increased in tumor samples compared with control skin. The concentration of VEGF-A was significantly increased within melanoma compared with control (L). Results of 9 different melanoma patients are shown (* P

    Journal: Blood

    Article Title: von Willebrand factor fibers promote cancer-associated platelet aggregation in malignant melanoma of mice and humans

    doi: 10.1182/blood-2014-08-595686

    Figure Lengend Snippet: Local inhibition of ADAMTS13 promotes VWF fiber formation in microvessels obtained from human melanoma patients. Cryosections of human malignant melanoma tissue, healthy control skin, and basal cell carcinoma were analyzed by immunofluorescence stainings for VWF and CD31 (A-C) or thrombospondin (TSP) (D-E). Nuclei were stained with DAPI. Analysis of healthy skin (A) and human basal cell carcinoma (B) as control demonstrate storage of VWF in the vessel wall (arrowheads). ULVWF fibers are detected in the lumen of the microvessels, correlating to reduced VWF within the vessel wall indicative of EC activation (C, arrows). These ULVWF fibers bind platelets (D, arrows) and are associated with microthrombi formation in distinct microvessels (E, asterisk; n = 5 to 6; scale bars = 20 µm). Quantification showed significantly increased numbers of vessels with luminal VWF fibers in tumor vasculature compared with healthy skin (F). Systemic VWF level in blood samples of malignant melanoma patients was increased compared with healthy control (G). By contrast, only a slight reduction of ADAMTS13 activity was observed (H). Tumor-derived cytokines and growth factors were measured in healthy control skin and tumor of human malignant melanoma by bio-plex. Cytokine levels of IFN-γ (I), TNF-α (J), and IL-6 (K) were increased in tumor samples compared with control skin. The concentration of VEGF-A was significantly increased within melanoma compared with control (L). Results of 9 different melanoma patients are shown (* P

    Article Snippet: Cryosections (10 µm) were incubated with the following primary antibodies: rabbit anti-human VWF (DakoCytomation), rat anti-mouse CD42b (emfret Analytics), mouse anti-human Thrombospondin (Laboratory Vision/Neomarkers), mouse anti-human CD31 (DakoCytomation), rat anti-mouse CD31 (BD Biosciences), Ki67–fluorescein isothiocyanate (FITC) (BD Biosciences), rabbit anti-VEGF-A (Santa Cruz Biotechnology).

    Techniques: Inhibition, Immunofluorescence, Staining, Activation Assay, Activity Assay, Derivative Assay, Concentration Assay

    Reduced ADAMTS13 activity in tumor tissue promotes intraluminal VWF fiber formation in tumor microvessels. Immunofluorescence staining of cryosections for VWF (green), platelets (red), and the endothelial cell marker CD31 (red). DNA was stained with DAPI (blue). Mouse melanoma cells (Ret) were injected intradermally and mice were treated with recombinant ADAMTS13 (rADAMTS13; E-F) or 0.9% NaCl (C-D) as control. Reconstitution of ADAMTS13 reduced the formation of intraluminal ULVWF networks and platelet aggregation compared with vehicle treatment. In comparison with wild-type skin (A-B), less VWF in the vessel wall indicates ULVWF degradation after exocytosis (n = 5-6 animals; scale bars = 20 µm). To analyze the impact of ADAMTS13 on VWF fiber formation, vessels with or without luminal fibers were quantified. Infusion with rADAMTS13 significantly decreased the number of vessels with ULVWF (G). The activity and the protein expression of ADAMTS13 in tumor tissue were significantly reduced compared with healthy skin measured by a FRET-based assay and western blot (H; n = 3-5 animals, * P

    Journal: Blood

    Article Title: von Willebrand factor fibers promote cancer-associated platelet aggregation in malignant melanoma of mice and humans

    doi: 10.1182/blood-2014-08-595686

    Figure Lengend Snippet: Reduced ADAMTS13 activity in tumor tissue promotes intraluminal VWF fiber formation in tumor microvessels. Immunofluorescence staining of cryosections for VWF (green), platelets (red), and the endothelial cell marker CD31 (red). DNA was stained with DAPI (blue). Mouse melanoma cells (Ret) were injected intradermally and mice were treated with recombinant ADAMTS13 (rADAMTS13; E-F) or 0.9% NaCl (C-D) as control. Reconstitution of ADAMTS13 reduced the formation of intraluminal ULVWF networks and platelet aggregation compared with vehicle treatment. In comparison with wild-type skin (A-B), less VWF in the vessel wall indicates ULVWF degradation after exocytosis (n = 5-6 animals; scale bars = 20 µm). To analyze the impact of ADAMTS13 on VWF fiber formation, vessels with or without luminal fibers were quantified. Infusion with rADAMTS13 significantly decreased the number of vessels with ULVWF (G). The activity and the protein expression of ADAMTS13 in tumor tissue were significantly reduced compared with healthy skin measured by a FRET-based assay and western blot (H; n = 3-5 animals, * P

    Article Snippet: Cryosections (10 µm) were incubated with the following primary antibodies: rabbit anti-human VWF (DakoCytomation), rat anti-mouse CD42b (emfret Analytics), mouse anti-human Thrombospondin (Laboratory Vision/Neomarkers), mouse anti-human CD31 (DakoCytomation), rat anti-mouse CD31 (BD Biosciences), Ki67–fluorescein isothiocyanate (FITC) (BD Biosciences), rabbit anti-VEGF-A (Santa Cruz Biotechnology).

    Techniques: Activity Assay, Immunofluorescence, Staining, Marker, Injection, Mouse Assay, Recombinant, Expressing, Western Blot

    Tinzaparin attenuates proliferation of melanoma cells in primary tumor and lymph nodes and attenuates tumor progression in ret transgenic mice. Immunofluorescence stainings of tumor cell proliferation with Ki67 (green) and DAPI (nuclei, blue). Cryosections of vehicle-treated tumors (A) show more proliferating melanoma cells compared with tinzaparin-treated tumors (B; scale bars = 50 µm). Quantification showed a significant reduction of Ki67-positive cells after treatment with tinzaparin (C). Analysis of Ki67-positive tumor cells in lymph nodes of vehicle-treated ret mice and tinzaparin-treated animals was assessed by flow cytometry. Results show that tinzaparin induces a significant reduction of proliferating melanoma cells (D). Bars show mean ± SD (n = 7-10). Tumor-derived VEGF was measured in tumor and lymph nodes of ret transgenic mice (E-F) by bio-plex assay. VEGF levels were decreased in tumor samples (E) and lymph nodes (F) after treatment with tinzaparin (n = 5-7 different mice each group). Consequently, a survival benefit (G) and a reduced tumor weight (H) compared with vehicle-treated control was observed after tinzaparin treatment (n = 13 of 2 independent experiments). Plot shows mean ± SD; * P

    Journal: Blood

    Article Title: von Willebrand factor fibers promote cancer-associated platelet aggregation in malignant melanoma of mice and humans

    doi: 10.1182/blood-2014-08-595686

    Figure Lengend Snippet: Tinzaparin attenuates proliferation of melanoma cells in primary tumor and lymph nodes and attenuates tumor progression in ret transgenic mice. Immunofluorescence stainings of tumor cell proliferation with Ki67 (green) and DAPI (nuclei, blue). Cryosections of vehicle-treated tumors (A) show more proliferating melanoma cells compared with tinzaparin-treated tumors (B; scale bars = 50 µm). Quantification showed a significant reduction of Ki67-positive cells after treatment with tinzaparin (C). Analysis of Ki67-positive tumor cells in lymph nodes of vehicle-treated ret mice and tinzaparin-treated animals was assessed by flow cytometry. Results show that tinzaparin induces a significant reduction of proliferating melanoma cells (D). Bars show mean ± SD (n = 7-10). Tumor-derived VEGF was measured in tumor and lymph nodes of ret transgenic mice (E-F) by bio-plex assay. VEGF levels were decreased in tumor samples (E) and lymph nodes (F) after treatment with tinzaparin (n = 5-7 different mice each group). Consequently, a survival benefit (G) and a reduced tumor weight (H) compared with vehicle-treated control was observed after tinzaparin treatment (n = 13 of 2 independent experiments). Plot shows mean ± SD; * P

    Article Snippet: Cryosections (10 µm) were incubated with the following primary antibodies: rabbit anti-human VWF (DakoCytomation), rat anti-mouse CD42b (emfret Analytics), mouse anti-human Thrombospondin (Laboratory Vision/Neomarkers), mouse anti-human CD31 (DakoCytomation), rat anti-mouse CD31 (BD Biosciences), Ki67–fluorescein isothiocyanate (FITC) (BD Biosciences), rabbit anti-VEGF-A (Santa Cruz Biotechnology).

    Techniques: Transgenic Assay, Mouse Assay, Immunofluorescence, Flow Cytometry, Cytometry, Derivative Assay, Plex Assay

    Immunofluorescence analysis of tumor microvessels compared with healthy control skin in ret transgenic mice. Cryosections were stained for VWF and CD31 (A,C). An anti-GPIb antibody was used to identify platelets (B,D). Nuclei were stained with DAPI. Representative images of control skin show VWF localized within the vessel wall, lacking ULVWF fibers within the lumen (A-B, arrowheads) and only few platelets are visible (B, red). By contrast, in tumor microvessels, ULVWF fibers are detectable within the vessel lumen indicating EC activation (C, arrows). These ULVWF fibers bind platelets as shown in the same vessel (D, red, arrows). Insets, A higher magnification of the presented images (n = 4 to 10 animals; scale bars = 20 µm). See also supplemental Figures 1-2.

    Journal: Blood

    Article Title: von Willebrand factor fibers promote cancer-associated platelet aggregation in malignant melanoma of mice and humans

    doi: 10.1182/blood-2014-08-595686

    Figure Lengend Snippet: Immunofluorescence analysis of tumor microvessels compared with healthy control skin in ret transgenic mice. Cryosections were stained for VWF and CD31 (A,C). An anti-GPIb antibody was used to identify platelets (B,D). Nuclei were stained with DAPI. Representative images of control skin show VWF localized within the vessel wall, lacking ULVWF fibers within the lumen (A-B, arrowheads) and only few platelets are visible (B, red). By contrast, in tumor microvessels, ULVWF fibers are detectable within the vessel lumen indicating EC activation (C, arrows). These ULVWF fibers bind platelets as shown in the same vessel (D, red, arrows). Insets, A higher magnification of the presented images (n = 4 to 10 animals; scale bars = 20 µm). See also supplemental Figures 1-2.

    Article Snippet: Cryosections (10 µm) were incubated with the following primary antibodies: rabbit anti-human VWF (DakoCytomation), rat anti-mouse CD42b (emfret Analytics), mouse anti-human Thrombospondin (Laboratory Vision/Neomarkers), mouse anti-human CD31 (DakoCytomation), rat anti-mouse CD31 (BD Biosciences), Ki67–fluorescein isothiocyanate (FITC) (BD Biosciences), rabbit anti-VEGF-A (Santa Cruz Biotechnology).

    Techniques: Immunofluorescence, Transgenic Assay, Mouse Assay, Staining, Activation Assay

    CCN1 inhibits hepatic fibrosis by inducing myofibroblast senescence. (A) Adjacent serial liver cryosections from Ccn1 flox / flox and Ccn1 Δ Hep mice treated with CCl 4 to induce fibrosis were stained for SA-β-Gal (upper panels, blue; n = 7)

    Journal: Molecular and Cellular Biology

    Article Title: Matricellular Protein CCN1 Promotes Regression of Liver Fibrosis through Induction of Cellular Senescence in Hepatic Myofibroblasts

    doi: 10.1128/MCB.00049-13

    Figure Lengend Snippet: CCN1 inhibits hepatic fibrosis by inducing myofibroblast senescence. (A) Adjacent serial liver cryosections from Ccn1 flox / flox and Ccn1 Δ Hep mice treated with CCl 4 to induce fibrosis were stained for SA-β-Gal (upper panels, blue; n = 7)

    Article Snippet: Cell proliferation in cryosections was assessed by immunohistochemistry using antibodies against Ki67 (Abcam).

    Techniques: Mouse Assay, Staining

    Sequential Expression of Germ-Cell-Related Transcription Factors in Single Cells during hPGCLC Specification (A and B) Immunofluorscence analysis for (A) BLIMP1, SOX17, and TFAP2C and (B) BLIMP1 and T in cryosections of day 1–8 embryoids after hPGCLC induction. Bottom row in (B) shows high exposure (digital) image of T, indicating low but specific expression in hPGCLC. SOX17-positive or BLIMP1-positive cells are highlighted. Scale bars, 50 μm. (C) Percentage of SOX17-positive (+) cells in day 1–8 embryoids that were also TFAP2C+ or BLIMP1+. Corresponds to data in Figure 4 A. (D) Percentage of BLIMP1-positive (+) cells in day 1–8 embryoids that were TFAP2C+, NANOG+, or OCT4+. Corresponds to data in Figures 4 A, S4 A, and S4B. (E) Summary model for dynamics of hPGCLC specification in embryoids. SOX17-positive cells are first scattered in day 1 embryoids. They gain expression of BLIMP1, TFAP2C, and NANOG sequentially and form a cluster from day 2 onward until the formation of nascent hPGCLC.

    Journal: Cell

    Article Title: SOX17 Is a Critical Specifier of Human Primordial Germ Cell Fate

    doi: 10.1016/j.cell.2014.12.013

    Figure Lengend Snippet: Sequential Expression of Germ-Cell-Related Transcription Factors in Single Cells during hPGCLC Specification (A and B) Immunofluorscence analysis for (A) BLIMP1, SOX17, and TFAP2C and (B) BLIMP1 and T in cryosections of day 1–8 embryoids after hPGCLC induction. Bottom row in (B) shows high exposure (digital) image of T, indicating low but specific expression in hPGCLC. SOX17-positive or BLIMP1-positive cells are highlighted. Scale bars, 50 μm. (C) Percentage of SOX17-positive (+) cells in day 1–8 embryoids that were also TFAP2C+ or BLIMP1+. Corresponds to data in Figure 4 A. (D) Percentage of BLIMP1-positive (+) cells in day 1–8 embryoids that were TFAP2C+, NANOG+, or OCT4+. Corresponds to data in Figures 4 A, S4 A, and S4B. (E) Summary model for dynamics of hPGCLC specification in embryoids. SOX17-positive cells are first scattered in day 1 embryoids. They gain expression of BLIMP1, TFAP2C, and NANOG sequentially and form a cluster from day 2 onward until the formation of nascent hPGCLC.

    Article Snippet: Samples were prepared as 8 μm cryosections on Superfrost Plus Micro slides (VWR) by a cryostat (Leica, 3050S).

    Techniques: Expressing

    Role of BLIMP1 in hPGCLC Specification (A) Western blot analysis of BLIMP1 and SOX17 in TNAP-positive (TNAP+) cells sorted from wild-type (WT) and BLIMP1 knockout (BLIMP1 KO) day 4 embryoids after hPGCLC induction. TUBULIN was used as loading control. (B) FACS analysis of TNAP and NANOS3-mCherry on WT and BLIMP1 knockout (BLIMP1 KO) day 4 embryoids. (C) Immunofluorscence for OCT4 and SOX17 in cryosections of WT and BLIMP1 KO day 4 and 8 embryoids. OCT4-positive cells are highlighted. Scale bar, 50 μm. (D) Expression analysis by RT-qPCR for WT TNAP/NANOS3-mCherry double-positive cells (WT; TNAP+N3+) and BLIMP1 KO TNAP single-positive cells (BLIMP1 KO; TNAP+) sorted from day 4 embryoids. Relative expression levels are shown with normalization to β−ACTIN . Error bars indicate mean ± SD from two independent biological replicates.

    Journal: Cell

    Article Title: SOX17 Is a Critical Specifier of Human Primordial Germ Cell Fate

    doi: 10.1016/j.cell.2014.12.013

    Figure Lengend Snippet: Role of BLIMP1 in hPGCLC Specification (A) Western blot analysis of BLIMP1 and SOX17 in TNAP-positive (TNAP+) cells sorted from wild-type (WT) and BLIMP1 knockout (BLIMP1 KO) day 4 embryoids after hPGCLC induction. TUBULIN was used as loading control. (B) FACS analysis of TNAP and NANOS3-mCherry on WT and BLIMP1 knockout (BLIMP1 KO) day 4 embryoids. (C) Immunofluorscence for OCT4 and SOX17 in cryosections of WT and BLIMP1 KO day 4 and 8 embryoids. OCT4-positive cells are highlighted. Scale bar, 50 μm. (D) Expression analysis by RT-qPCR for WT TNAP/NANOS3-mCherry double-positive cells (WT; TNAP+N3+) and BLIMP1 KO TNAP single-positive cells (BLIMP1 KO; TNAP+) sorted from day 4 embryoids. Relative expression levels are shown with normalization to β−ACTIN . Error bars indicate mean ± SD from two independent biological replicates.

    Article Snippet: Samples were prepared as 8 μm cryosections on Superfrost Plus Micro slides (VWR) by a cryostat (Leica, 3050S).

    Techniques: Western Blot, Knock-Out, FACS, Expressing, Quantitative RT-PCR

    Sequential Expression of Germ-Cell-Related Genes during PGCLC Specification, Related to Figure 4 (A–C) Immunofluoresence of (A) PRDM14 and NANOG; (B) OCT4; and (C) NANOS3-mCherry on cryosections of day 1-8. hPGCLC were counterstained with BLIMP1 or OCT4 as highlighted. Arrowheads indicate enrichment of PRDM14 in cytoplasm. Scale bars = 70 μm.

    Journal: Cell

    Article Title: SOX17 Is a Critical Specifier of Human Primordial Germ Cell Fate

    doi: 10.1016/j.cell.2014.12.013

    Figure Lengend Snippet: Sequential Expression of Germ-Cell-Related Genes during PGCLC Specification, Related to Figure 4 (A–C) Immunofluoresence of (A) PRDM14 and NANOG; (B) OCT4; and (C) NANOS3-mCherry on cryosections of day 1-8. hPGCLC were counterstained with BLIMP1 or OCT4 as highlighted. Arrowheads indicate enrichment of PRDM14 in cytoplasm. Scale bars = 70 μm.

    Article Snippet: Samples were prepared as 8 μm cryosections on Superfrost Plus Micro slides (VWR) by a cryostat (Leica, 3050S).

    Techniques: Expressing

    Immunofluorescence for Epigenetic Modifications and Modifiers in Embryoids, Related to Figure 3 (A–C) Immunofluorescence analysis was carried out for (A) 5-methylcytosine (5mC); (B) DNMT3A; (C) UHRF1 on day 4 embryoids. TFAP2C, OCT4 or BLIMP1 were used to counterstain for hPGCLCs. hPGCLCs were highlighted by white dashed lines. White arrowheads showed examples of KI-67-positive PGCLCs while yellow arrows showed the KI-67-negative PGCLCs (C). Scale bars = 50 μm. (D) Expression level of methylation related epigenetic modifiers from RNA-Seq dataset. Mean normalized read counts from two biological replicates were shown. (E) Immunofluoresence of PRMT5 on cryosections of embryoids collected at day 1, 2, 4 and 8 post-hPGCLC induction. hPGCLCs were counterstained with OCT4 as highlighted. Scale bar = 70 μm.

    Journal: Cell

    Article Title: SOX17 Is a Critical Specifier of Human Primordial Germ Cell Fate

    doi: 10.1016/j.cell.2014.12.013

    Figure Lengend Snippet: Immunofluorescence for Epigenetic Modifications and Modifiers in Embryoids, Related to Figure 3 (A–C) Immunofluorescence analysis was carried out for (A) 5-methylcytosine (5mC); (B) DNMT3A; (C) UHRF1 on day 4 embryoids. TFAP2C, OCT4 or BLIMP1 were used to counterstain for hPGCLCs. hPGCLCs were highlighted by white dashed lines. White arrowheads showed examples of KI-67-positive PGCLCs while yellow arrows showed the KI-67-negative PGCLCs (C). Scale bars = 50 μm. (D) Expression level of methylation related epigenetic modifiers from RNA-Seq dataset. Mean normalized read counts from two biological replicates were shown. (E) Immunofluoresence of PRMT5 on cryosections of embryoids collected at day 1, 2, 4 and 8 post-hPGCLC induction. hPGCLCs were counterstained with OCT4 as highlighted. Scale bar = 70 μm.

    Article Snippet: Samples were prepared as 8 μm cryosections on Superfrost Plus Micro slides (VWR) by a cryostat (Leica, 3050S).

    Techniques: Immunofluorescence, Expressing, Methylation, RNA Sequencing Assay

    MAPC-engrafted mouse with chronic GVHD. Intravital microscopy and histopathological findings in one NOD-SCID mouse that received 0.6 × 10 6 MAPCs 16 wk earlier are consistent with the development of chronic GVHD. Left panels show the presence of GFP + cells in the GVHD target organs, specifically skin (underlayer), liver, lung, and ileum. All intravital images were taken at a zoom factor of 8× and a transfer lens of 0.63× with an MZFLIII stereomicroscope (300 millisecond exposure). Right panels show the corresponding hematoxylin and eosin stains of cryosections taken from the same mouse (bar, 900 μm). The skin shows inflammation in the dermis and subdermis (black arrow), extensive epidermal hyperplasia (white arrow), and dyskeratosis (asterisk). GVHD score, 3.5 (0–4 scale). The liver has moderate inflammation around the portal triads with evidence of bile duct degeneration (black arrow). GVHD score, 3.0. The lung has peribronchiolar inflammation (b, bronchiole) and extensive parenchymal inflammation in surrounding areas with organizing alveolitis and fibrosis. GVHD score, 4.0. The ileum shows only mild inflammation of the lamina propria as is typical for chronic GVHD. GVHD score, 0.5.

    Journal: The Journal of Experimental Medicine

    Article Title: Hematopoietic reconstitution by multipotent adult progenitor cells: precursors to long-term hematopoietic stem cells

    doi: 10.1084/jem.20061115

    Figure Lengend Snippet: MAPC-engrafted mouse with chronic GVHD. Intravital microscopy and histopathological findings in one NOD-SCID mouse that received 0.6 × 10 6 MAPCs 16 wk earlier are consistent with the development of chronic GVHD. Left panels show the presence of GFP + cells in the GVHD target organs, specifically skin (underlayer), liver, lung, and ileum. All intravital images were taken at a zoom factor of 8× and a transfer lens of 0.63× with an MZFLIII stereomicroscope (300 millisecond exposure). Right panels show the corresponding hematoxylin and eosin stains of cryosections taken from the same mouse (bar, 900 μm). The skin shows inflammation in the dermis and subdermis (black arrow), extensive epidermal hyperplasia (white arrow), and dyskeratosis (asterisk). GVHD score, 3.5 (0–4 scale). The liver has moderate inflammation around the portal triads with evidence of bile duct degeneration (black arrow). GVHD score, 3.0. The lung has peribronchiolar inflammation (b, bronchiole) and extensive parenchymal inflammation in surrounding areas with organizing alveolitis and fibrosis. GVHD score, 4.0. The ileum shows only mild inflammation of the lamina propria as is typical for chronic GVHD. GVHD score, 0.5.

    Article Snippet: 6-μm cryosections were thaw-mounted onto glass slides and fixed in acetone for 5 min. Spleen, LNs, and thymus were immunostained for donor- derived lymphocytes using anti–CD3-PE, anti–CD4-biotin, rabbit anti–GFP, anti–CD19-PE, anti–CD11b-PE, anti–CD11c-biotin, and anti–CD45.2-biotin (all from BD Biosciences).

    Techniques: Intravital Microscopy

    Venules are the source of hemorrhage. A–D: Cryosections immunolabeled for laminin (brown) , which identifies both arterioles and venules, and stained for alkaline phosphatase (blue) , which labels only arterioles, showing hemorrhages in the cortex

    Journal: Journal of neurosurgery. Pediatrics

    Article Title: Tandem insults of prenatal ischemia plus postnatal raised intrathoracic pressure in a novel rat model of encephalopathy of prematurity

    doi: 10.3171/2011.9.PEDS11174

    Figure Lengend Snippet: Venules are the source of hemorrhage. A–D: Cryosections immunolabeled for laminin (brown) , which identifies both arterioles and venules, and stained for alkaline phosphatase (blue) , which labels only arterioles, showing hemorrhages in the cortex

    Article Snippet: Cryosections (10 µm) were collected on slides, blocked in 5% goat serum with 0.2% Triton X-100 in PBS for 1 hour, then incubated overnight with primary antibodies directed against ED-1 (CD68) (dilution 1:500, MAB1435), cleaved caspase-3 (dilution 1:200, Cell Signaling Technology), or laminin (dilution 1:200; FA-2404–1, EY Laboratories, Inc.) at 4°C.

    Techniques: Immunolabeling, Staining

    Progressive thymic epithelium phenotypes in Foxn1 lacZ mutants . (A-D) Cryosections were stained for Keratin 8 (green) and Keratin 5 (red). Scale bar: 200 μm. Arrowhead, perivascular space; *, cystic structure. (A) Two-week + /lacZ , + /nu , and lacZ/lacZ

    Journal: Blood

    Article Title: Foxn1 is required to maintain the postnatal thymic microenvironment in a dosage-sensitive manner

    doi: 10.1182/blood-2008-05-156265

    Figure Lengend Snippet: Progressive thymic epithelium phenotypes in Foxn1 lacZ mutants . (A-D) Cryosections were stained for Keratin 8 (green) and Keratin 5 (red). Scale bar: 200 μm. Arrowhead, perivascular space; *, cystic structure. (A) Two-week + /lacZ , + /nu , and lacZ/lacZ

    Article Snippet: Immunofluorescence was performed on cryosections using the following antibodies: rabbit anti–mouse K5, rabbit anti–mouse K14 (Covance Research Products, Princeton, NJ), rat anti–mouse K8 clone (Troma-1; Developmental Studies Hybridoma Bank, Iowa City, IA), or with UEA-1–biotin (Vector Laboratories, Burlingame, CA), donkey anti–rat IgG-FITC, donkey anti–rabbit IgG–Texas Red secondary antibodies, streptavidin-FITC (Jackson Immunoresearch Laboratories, West Grove, PA), goat anti–mouse Foxn1 (WHN G-20; Santa Cruz Biotechnology, Santa Cruz, CA), and rat anti–mouse MHCII (BD Pharmingen, San Diego, CA).

    Techniques: Staining

    Oral treatment with Ac 4 ManNAc prevents appearance of rimmed vacuoles. A , histochemistry of gastrocnemius cryosections from DMRV/hIBM mice in treated and placebo groups is shown. In the placebo group, H E staining shows variations in fiber size,

    Journal: The Journal of Biological Chemistry

    Article Title: Peracetylated N-Acetylmannosamine, a Synthetic Sugar Molecule, Efficiently Rescues Muscle Phenotype and Biochemical Defects in Mouse Model of Sialic Acid-deficient Myopathy *

    doi: 10.1074/jbc.M111.297051

    Figure Lengend Snippet: Oral treatment with Ac 4 ManNAc prevents appearance of rimmed vacuoles. A , histochemistry of gastrocnemius cryosections from DMRV/hIBM mice in treated and placebo groups is shown. In the placebo group, H E staining shows variations in fiber size,

    Article Snippet: We immunostained 6-μm-thick cryosections from gastrocnemius muscles using the primary Abs rat mAb to lysosome-associated membrane protein 2 (Lamp2) (clone ABL-93, Developmental Studies Hybridoma Bank), rabbit polyclonal Ab to Aβ1–42 (AB5078P, Millipore, Billerica, MA), and mAb to polyubiquitin (Enzo Life Sciences, Inc. Farmingdale, NY) following published protocols ( , ).

    Techniques: Mouse Assay, Staining

    Oral treatment with Ac 4 ManNAc prevents occurrence of intracellular inclusions in DMRV/hIBM myofibers. A , in muscle cryosections from non-treated DMRV/hIBM mice (DMRV placebo), immunoreactive signals to Lamp2, Aβ1–42, and polyubiquitin

    Journal: The Journal of Biological Chemistry

    Article Title: Peracetylated N-Acetylmannosamine, a Synthetic Sugar Molecule, Efficiently Rescues Muscle Phenotype and Biochemical Defects in Mouse Model of Sialic Acid-deficient Myopathy *

    doi: 10.1074/jbc.M111.297051

    Figure Lengend Snippet: Oral treatment with Ac 4 ManNAc prevents occurrence of intracellular inclusions in DMRV/hIBM myofibers. A , in muscle cryosections from non-treated DMRV/hIBM mice (DMRV placebo), immunoreactive signals to Lamp2, Aβ1–42, and polyubiquitin

    Article Snippet: We immunostained 6-μm-thick cryosections from gastrocnemius muscles using the primary Abs rat mAb to lysosome-associated membrane protein 2 (Lamp2) (clone ABL-93, Developmental Studies Hybridoma Bank), rabbit polyclonal Ab to Aβ1–42 (AB5078P, Millipore, Billerica, MA), and mAb to polyubiquitin (Enzo Life Sciences, Inc. Farmingdale, NY) following published protocols ( , ).

    Techniques: Mouse Assay