Structured Review

The Jackson Laboratory wt c57bl 6j mice
ProBDNF protein levels are elevated acutely after pilocarpine-induced SE in WT <t>C57BL/6J</t> mice. A , Bottom, Representative Western blot of whole hippocampal protein homogenates from WT mice killed 3 h after the induction of SE or time-matched saline controls probed with proBDNF (1:1000) and anti-actin antibodies. Top, Densitometry analysis of proBDNF protein abundance. Ratio of proBDNF/actin at 3 h after SE ( N = 5), expressed as the percentage change relative to mean values (±SEM) of the control group ( N = 5; **** p
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1) Product Images from "Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2 3"

Article Title: Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2 3

Journal: eNeuro

doi: 10.1523/ENEURO.0020-15.2016

ProBDNF protein levels are elevated acutely after pilocarpine-induced SE in WT C57BL/6J mice. A , Bottom, Representative Western blot of whole hippocampal protein homogenates from WT mice killed 3 h after the induction of SE or time-matched saline controls probed with proBDNF (1:1000) and anti-actin antibodies. Top, Densitometry analysis of proBDNF protein abundance. Ratio of proBDNF/actin at 3 h after SE ( N = 5), expressed as the percentage change relative to mean values (±SEM) of the control group ( N = 5; **** p
Figure Legend Snippet: ProBDNF protein levels are elevated acutely after pilocarpine-induced SE in WT C57BL/6J mice. A , Bottom, Representative Western blot of whole hippocampal protein homogenates from WT mice killed 3 h after the induction of SE or time-matched saline controls probed with proBDNF (1:1000) and anti-actin antibodies. Top, Densitometry analysis of proBDNF protein abundance. Ratio of proBDNF/actin at 3 h after SE ( N = 5), expressed as the percentage change relative to mean values (±SEM) of the control group ( N = 5; **** p

Techniques Used: Mouse Assay, Western Blot

2) Product Images from "Fever Promotes T Lymphocyte Trafficking via a Thermal Sensory Pathway Involving Heat Shock Protein 90 and α4 Integrins"

Article Title: Fever Promotes T Lymphocyte Trafficking via a Thermal Sensory Pathway Involving Heat Shock Protein 90 and α4 Integrins

Journal: Immunity

doi: 10.1016/j.immuni.2018.11.013

Disruption of Hsp90-α4 Interaction Inhibits Fever-Induced T Cell Trafficking In vivo and Impairs the Clearance of Bacterial Infection (A–D) WT and KI C57BL/6J mice were treated with normothermia (NT; core temperature 36.8°C ± 0.2°C) or fever-range whole-body hyperthermia (WBH; core temperature 39.5°C ± 0.5°C) for 6 hr (n = 7–10 mice per group) and then were sacrificed. T cells were isolated from the spleen. α4β7-VCAM-1 binding was disrupted by cell pre-treatment with 10 μg/mL α4β7-blocking antibody DATK32 during examination of α4β1-mediated cell adhesion and migration on VCAM-1 substrate in (B) and (C). Co-immunoprecipitation of Hsp90AA1 and Hsp90AB1 with integrin α4 in the cell-membrane fractions is shown in (A). Adhesion of T cells to immobilized VCAM-1-Fc (5 μg/mL) or MAdCAM-1-Fc (5 μg/mL) substrate in 1 mM Ca 2+ + Mg 2+ at a wall shear stress of 1 dyn/cm 2 is shown in (B). Transmigration of T cells across membranes coated in VCAM-1-Fc (5 μg/mL) or MAdCAM-1-Fc (5 μg/mL) in the presence of CCL21 (500 ng/mL) in the lower chamber is shown in (C). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified (D). (E–G) WT and KI mice were injected with LPS (10 μg/kg) or PBS at time zero (n = 3 mice per group). Body temperature was monitored every hour. The bar shows the dark period (E). Immunoblot analysis of Hsp90AA1 and Hsp90AB1 in T cells isolated from PLNs in mice is shown in (F). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified (G). (H) Effect of different temperatures on the expression of Hsp90. T cells were isolated from PLNs in WT mice and then treated at 37°C, 38°C, 38.5°C, 39°C, or 40°C for 12 hr. Immunoblot analysis of Hsp90AA1 and Hsp90AB1 in T cells is shown. (I–M) WT and KI mice were orally injected with PBS or S. typhimurium strain SL1344 (10 8 CFU per mouse; n = 4–12 mice per group). Body temperature was monitored for 5 days (I). The survival rates of WT and KI mice are shown in (J), and significance was calculated as given. H E staining of the small intestine at day 5 after infection is shown in (K) (scale bar, 100 mm). Immunofluorescence analysis of the small intestine sections at day 5 after infection is shown. Stain coloring is as follows: DAPI, blue; S. typhimurium expressing GFP, green; and CD3, red. Quantifications of S. typhimurium colonies and CD3 + cells are shown below in (L) (scale bar, 100 mm). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified at day 5 after infection (M). .
Figure Legend Snippet: Disruption of Hsp90-α4 Interaction Inhibits Fever-Induced T Cell Trafficking In vivo and Impairs the Clearance of Bacterial Infection (A–D) WT and KI C57BL/6J mice were treated with normothermia (NT; core temperature 36.8°C ± 0.2°C) or fever-range whole-body hyperthermia (WBH; core temperature 39.5°C ± 0.5°C) for 6 hr (n = 7–10 mice per group) and then were sacrificed. T cells were isolated from the spleen. α4β7-VCAM-1 binding was disrupted by cell pre-treatment with 10 μg/mL α4β7-blocking antibody DATK32 during examination of α4β1-mediated cell adhesion and migration on VCAM-1 substrate in (B) and (C). Co-immunoprecipitation of Hsp90AA1 and Hsp90AB1 with integrin α4 in the cell-membrane fractions is shown in (A). Adhesion of T cells to immobilized VCAM-1-Fc (5 μg/mL) or MAdCAM-1-Fc (5 μg/mL) substrate in 1 mM Ca 2+ + Mg 2+ at a wall shear stress of 1 dyn/cm 2 is shown in (B). Transmigration of T cells across membranes coated in VCAM-1-Fc (5 μg/mL) or MAdCAM-1-Fc (5 μg/mL) in the presence of CCL21 (500 ng/mL) in the lower chamber is shown in (C). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified (D). (E–G) WT and KI mice were injected with LPS (10 μg/kg) or PBS at time zero (n = 3 mice per group). Body temperature was monitored every hour. The bar shows the dark period (E). Immunoblot analysis of Hsp90AA1 and Hsp90AB1 in T cells isolated from PLNs in mice is shown in (F). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified (G). (H) Effect of different temperatures on the expression of Hsp90. T cells were isolated from PLNs in WT mice and then treated at 37°C, 38°C, 38.5°C, 39°C, or 40°C for 12 hr. Immunoblot analysis of Hsp90AA1 and Hsp90AB1 in T cells is shown. (I–M) WT and KI mice were orally injected with PBS or S. typhimurium strain SL1344 (10 8 CFU per mouse; n = 4–12 mice per group). Body temperature was monitored for 5 days (I). The survival rates of WT and KI mice are shown in (J), and significance was calculated as given. H E staining of the small intestine at day 5 after infection is shown in (K) (scale bar, 100 mm). Immunofluorescence analysis of the small intestine sections at day 5 after infection is shown. Stain coloring is as follows: DAPI, blue; S. typhimurium expressing GFP, green; and CD3, red. Quantifications of S. typhimurium colonies and CD3 + cells are shown below in (L) (scale bar, 100 mm). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified at day 5 after infection (M). .

Techniques Used: In Vivo, Infection, Mouse Assay, Isolation, Binding Assay, Blocking Assay, Migration, Immunoprecipitation, Transmigration Assay, Injection, Expressing, Staining, Immunofluorescence

3) Product Images from "Rickettsia parkeri Sca2 promotes dissemination in an intradermal infection mouse model"

Article Title: Rickettsia parkeri Sca2 promotes dissemination in an intradermal infection mouse model

Journal: bioRxiv

doi: 10.1101/2020.09.23.310409

R. parkeri Sca2 contributes to dissemination from skin to peripheral organs. a ) Survival of Ifnar −/− Ifngr −/− mice upon i.v. infection with 5 × 10 6 R. parkeri . n =7 (WT), 10 ( sca2 ::Tn), and 7 ( MC1_RS08740 ::Tn R. parkeri ) individual mice. Data are the combination of two independent experiments. b ) Survival of Ifnar −/− Ifngr −/− mice upon i.v. infection with 10 7 R. parkeri . n =7 (WT) and 10 ( sca2 ::Tn) individual mice. Data are the combination of two independent experiments. c ) Survival of Ifnar −/− Ifngr −/− mice upon i.d. infection with 10 7 R. parkeri . n =6 (WT) and 8 ( sca2 ::Tn) individual mice. Data are the combination of two independent experiments. d ) Weight changes of Ifnar −/− Ifngr −/− mice upon i.d. infection with 10 7 R. parkeri . n =6 (WT) and 8 ( sca2 ::Tn) individual mice. Data are the combination of two independent experiments. e ) Analysis of gross skin pathology after i.d. infection. Ifnar −/− Ifngr −/− mice were infected with 10 7 of the indicated strains of R. parkeri and monitored over time. n =9 (WT) and 8 ( sca2 ::Tn) individual mice. Data are the combination of two independent experiments. f ) Representative images of fluorescence in mouse skin after i.d. infection with 10 6 R. parkeri and delivery of a fluorescent dextran, at 5 d.p.i. Scale bars, 1 cm. The larger black dashed circle represents the area that was measured for fluorescence for each sample, as indicated in Fig. 3g (~80,000 pixels). The smaller black-dashed circle represents of the injection site area that was measured for fluorescence for each sample, as indicated in Extended Data Fig. 3 (~7,800 pixels). g ) Quantification of fluorescence in mouse skin after i.d. infection. Mice were infected with 10 7 R. parkeri , and 150 μl fluorescent dextran was intravenously delivered at 5 d.p.i. Skin was harvested 2 h later, and fluorescence was measured using a fluorescence imager. Data indicate measurements of larger areas of skin, as indicated in f by the larger black circle. n =6 (WT R. parkeri ) and n =6 ( sca2 ::Tn R. parkeri ) individual Ifnar −/− Ifngr −/− mice; n =6 (WT R. parkeri ) individual WT mice. For each experiment, the average of uninfected samples was normalized to 100; each sample was divided by the average for uninfected mice and multiplied by 100; the dotted horizontal line indicates 100 arbitrary units, corresponding to uninfected (unin.) mice. Data are the combination of two independent experiments. h ) Quantification of L. monocytogenes abundance in organs of WT C57BL/6J mice upon i.v. infection with 10 4 bacteria, at 72 h.p.i. Data are the combination of two independent experiments. n=6 (WT), n=7 (Δ actA ) individual mice. i ) Quantification of R. parkeri abundance in spleens and livers of WT C57BL/6J mice upon i.v. infection, at 72 h.p.i. Data are the combination of two independent experiments. n =10 (WT) and 10 ( Sca2 ::Tn) individual mice. j ) Quantification of R. parkeri abundance in organs upon i.d. infection with 10 7 R. parkeri . n =7 (WT) and 7 ( sca2 ::Tn) individual mice. Data are the combination of two independent experiments. Data for WT R. parkeri in Ifnar −/− Ifngr −/− mice are the same as in Fig. 2e . k ) Quantification of R. parkeri abundance in organs upon i.d. infection with 10 5 R. parkeri . n =7 (WT) and 6 ( sca2 ::Tn). Data are the combination of two independent experiments. Solid horizontal bars in g indicate means; solid horizontal bars in h-k indicate medians; error bars indicate SD. Statistical analyses for survival in a, b, c used a log-rank (Mantel-Cox) test. Statistical analysis in d used a two-way ANOVA at t=20. Statistical analysis in e used a two-way ANOVA from 0 to 10 d.p.i. Statistical analyses in g used a two-tailed Student’s T test. Statistical analyses in h, i, j, k used a two-tailed Mann-Whitney U test. The fold change in h, i, j, k indicates differences of medians. * P
Figure Legend Snippet: R. parkeri Sca2 contributes to dissemination from skin to peripheral organs. a ) Survival of Ifnar −/− Ifngr −/− mice upon i.v. infection with 5 × 10 6 R. parkeri . n =7 (WT), 10 ( sca2 ::Tn), and 7 ( MC1_RS08740 ::Tn R. parkeri ) individual mice. Data are the combination of two independent experiments. b ) Survival of Ifnar −/− Ifngr −/− mice upon i.v. infection with 10 7 R. parkeri . n =7 (WT) and 10 ( sca2 ::Tn) individual mice. Data are the combination of two independent experiments. c ) Survival of Ifnar −/− Ifngr −/− mice upon i.d. infection with 10 7 R. parkeri . n =6 (WT) and 8 ( sca2 ::Tn) individual mice. Data are the combination of two independent experiments. d ) Weight changes of Ifnar −/− Ifngr −/− mice upon i.d. infection with 10 7 R. parkeri . n =6 (WT) and 8 ( sca2 ::Tn) individual mice. Data are the combination of two independent experiments. e ) Analysis of gross skin pathology after i.d. infection. Ifnar −/− Ifngr −/− mice were infected with 10 7 of the indicated strains of R. parkeri and monitored over time. n =9 (WT) and 8 ( sca2 ::Tn) individual mice. Data are the combination of two independent experiments. f ) Representative images of fluorescence in mouse skin after i.d. infection with 10 6 R. parkeri and delivery of a fluorescent dextran, at 5 d.p.i. Scale bars, 1 cm. The larger black dashed circle represents the area that was measured for fluorescence for each sample, as indicated in Fig. 3g (~80,000 pixels). The smaller black-dashed circle represents of the injection site area that was measured for fluorescence for each sample, as indicated in Extended Data Fig. 3 (~7,800 pixels). g ) Quantification of fluorescence in mouse skin after i.d. infection. Mice were infected with 10 7 R. parkeri , and 150 μl fluorescent dextran was intravenously delivered at 5 d.p.i. Skin was harvested 2 h later, and fluorescence was measured using a fluorescence imager. Data indicate measurements of larger areas of skin, as indicated in f by the larger black circle. n =6 (WT R. parkeri ) and n =6 ( sca2 ::Tn R. parkeri ) individual Ifnar −/− Ifngr −/− mice; n =6 (WT R. parkeri ) individual WT mice. For each experiment, the average of uninfected samples was normalized to 100; each sample was divided by the average for uninfected mice and multiplied by 100; the dotted horizontal line indicates 100 arbitrary units, corresponding to uninfected (unin.) mice. Data are the combination of two independent experiments. h ) Quantification of L. monocytogenes abundance in organs of WT C57BL/6J mice upon i.v. infection with 10 4 bacteria, at 72 h.p.i. Data are the combination of two independent experiments. n=6 (WT), n=7 (Δ actA ) individual mice. i ) Quantification of R. parkeri abundance in spleens and livers of WT C57BL/6J mice upon i.v. infection, at 72 h.p.i. Data are the combination of two independent experiments. n =10 (WT) and 10 ( Sca2 ::Tn) individual mice. j ) Quantification of R. parkeri abundance in organs upon i.d. infection with 10 7 R. parkeri . n =7 (WT) and 7 ( sca2 ::Tn) individual mice. Data are the combination of two independent experiments. Data for WT R. parkeri in Ifnar −/− Ifngr −/− mice are the same as in Fig. 2e . k ) Quantification of R. parkeri abundance in organs upon i.d. infection with 10 5 R. parkeri . n =7 (WT) and 6 ( sca2 ::Tn). Data are the combination of two independent experiments. Solid horizontal bars in g indicate means; solid horizontal bars in h-k indicate medians; error bars indicate SD. Statistical analyses for survival in a, b, c used a log-rank (Mantel-Cox) test. Statistical analysis in d used a two-way ANOVA at t=20. Statistical analysis in e used a two-way ANOVA from 0 to 10 d.p.i. Statistical analyses in g used a two-tailed Student’s T test. Statistical analyses in h, i, j, k used a two-tailed Mann-Whitney U test. The fold change in h, i, j, k indicates differences of medians. * P

Techniques Used: Mouse Assay, Infection, Fluorescence, Injection, Two Tailed Test, MANN-WHITNEY

4) Product Images from "Consumptive coagulopathy of severe yellow fever occurs independently of hepatocellular tropism and massive hepatic injury"

Article Title: Consumptive coagulopathy of severe yellow fever occurs independently of hepatocellular tropism and massive hepatic injury

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.2014096117

YFV is viscerotropic in hFRG mice and macaques. ( A ) Weight-loss curves for macaques (mac), wild-type (WT) C57BL/6J mice (gray), muFRG mice (green), and hFRG mice (blue) following YFV inoculation. One macaque did not develop overt disease (denoted in salmon); this macaque was necropsied at day 18 post infection. ( B ) YFV viral load in plasma and ( C ) tissue homogenates, as determined by RT-qPCR. ( D ) Ratio of viral RNA load in brain versus liver, expressed as a ratio of the non–log-transformed value in each individual. ( E ) Quantification of apoptosis/necrosis in livers from YFV-infected macaques (red), muFRG mice (green), and hFRG mice (blue). Scoring of apoptosis/necrosis (and engraftment for hFRG mice, far right) was performed on hematoxylin- and eosin-stained sections by two board-certified hepatopathologists blinded to YFV-infection status. Data are the average of the two scores. ( F ) Linear regression of YFV burden in liver and plasma versus the percentage of liver scored as apoptosis/necrotic by examination of hematoxylin- and eosin-stained sections for YFV-infected macaques (red) and hFRG mice (blue). Statistical significance was determined from unpaired t tests performed on log-transformed values and is denoted by an open star (n.s.: P ≥ 0.05; ☆: P = 0.01 to 0.05; ☆☆☆☆: P
Figure Legend Snippet: YFV is viscerotropic in hFRG mice and macaques. ( A ) Weight-loss curves for macaques (mac), wild-type (WT) C57BL/6J mice (gray), muFRG mice (green), and hFRG mice (blue) following YFV inoculation. One macaque did not develop overt disease (denoted in salmon); this macaque was necropsied at day 18 post infection. ( B ) YFV viral load in plasma and ( C ) tissue homogenates, as determined by RT-qPCR. ( D ) Ratio of viral RNA load in brain versus liver, expressed as a ratio of the non–log-transformed value in each individual. ( E ) Quantification of apoptosis/necrosis in livers from YFV-infected macaques (red), muFRG mice (green), and hFRG mice (blue). Scoring of apoptosis/necrosis (and engraftment for hFRG mice, far right) was performed on hematoxylin- and eosin-stained sections by two board-certified hepatopathologists blinded to YFV-infection status. Data are the average of the two scores. ( F ) Linear regression of YFV burden in liver and plasma versus the percentage of liver scored as apoptosis/necrotic by examination of hematoxylin- and eosin-stained sections for YFV-infected macaques (red) and hFRG mice (blue). Statistical significance was determined from unpaired t tests performed on log-transformed values and is denoted by an open star (n.s.: P ≥ 0.05; ☆: P = 0.01 to 0.05; ☆☆☆☆: P

Techniques Used: Mouse Assay, Infection, Quantitative RT-PCR, Transformation Assay, Staining

5) Product Images from "Generation of Functional Blood Vessels from a Single c-kit+ Adult Vascular Endothelial Stem CellOn the Hunt for Vascular Endothelial Stem Cells"

Article Title: Generation of Functional Blood Vessels from a Single c-kit+ Adult Vascular Endothelial Stem CellOn the Hunt for Vascular Endothelial Stem Cells

Journal: PLoS Biology

doi: 10.1371/journal.pbio.1001407

A single c-kit-expressing adult VESC by the phenotype lin−CD31+CD105+Sca-1+CD117+ can generate functional, perfused blood vessels in vivo. (A) Flow diagram of the FACS sorting procedure used to obtain lin−CD31+CD105+Sca1+ CD117+ cells is shown. A single clonal colony originating from a single GFP-tagged lin−CD31+CD105+Sca1+CD117+ CFC was expanded for 12 d in adherent culture to amplify the cell number, manually picked up using a micropipette, mixed with 200 µl of matrigel supplemented with VEGF and bFGF, and injected subcutaneously into wt C57BL/6J mice. An eGFP channel inverted microscope micrograph of a single colony prior that was picked up and transplanted to a wt host is also shown. Scale bar, 150 µm. (B) Functional, perfused GFP+ blood vessels generated by the transplanted descendants of a single c-kit-expressing colony-forming EC by the phenotype lin− CD31+CD105+Sca1+CD117+ cell (14 d after transplantation). The mouse was perfused with fluorescent 0.2 µm microspheres (red) to stain endothelia in functional blood vessels that are connected to the blood circulation. ECs were stained for CD31 or CD105. Scale bars, 100 µm. 1-µm thick confocal optical slices and a 3-D orthogonal projection (x–z and y–z axes) are also shown. Note the blood vessel lumina (*) and the red endothelial signal from the microsphere perfusion of functional vasculature. Scale bars, 10 µm. Six independent experiments with similar results were performed. (C) Self-renewal capacity, a defining characteristic of stem cells, was evaluated by inoculating mice with syngeneic B16 melanomas (2 million cells per mice) together with 15 CFUs of GFP-tagged isolated CD31+CD105+ ECs. After 2 wk of tumor growth, repeated isolations and serial transplantations of lineage depleted single cell suspensions containing the GFP+ tagged ECs and the B16 cells were performed. The figure shows GFP+ blood vessels in the quaternary transplant. ECs were stained for VEGFR-2 (red), vWF (white), and CD31 and CD105 (red). Scale bar, 10 µm. A 3-D reconstitution of a GFP+ blood vessel in the quaternary transplant is also shown (right; a 34-µm thick stack of 34 x–y slices from a confocal scan). Six independent experiments with similar results were performed.
Figure Legend Snippet: A single c-kit-expressing adult VESC by the phenotype lin−CD31+CD105+Sca-1+CD117+ can generate functional, perfused blood vessels in vivo. (A) Flow diagram of the FACS sorting procedure used to obtain lin−CD31+CD105+Sca1+ CD117+ cells is shown. A single clonal colony originating from a single GFP-tagged lin−CD31+CD105+Sca1+CD117+ CFC was expanded for 12 d in adherent culture to amplify the cell number, manually picked up using a micropipette, mixed with 200 µl of matrigel supplemented with VEGF and bFGF, and injected subcutaneously into wt C57BL/6J mice. An eGFP channel inverted microscope micrograph of a single colony prior that was picked up and transplanted to a wt host is also shown. Scale bar, 150 µm. (B) Functional, perfused GFP+ blood vessels generated by the transplanted descendants of a single c-kit-expressing colony-forming EC by the phenotype lin− CD31+CD105+Sca1+CD117+ cell (14 d after transplantation). The mouse was perfused with fluorescent 0.2 µm microspheres (red) to stain endothelia in functional blood vessels that are connected to the blood circulation. ECs were stained for CD31 or CD105. Scale bars, 100 µm. 1-µm thick confocal optical slices and a 3-D orthogonal projection (x–z and y–z axes) are also shown. Note the blood vessel lumina (*) and the red endothelial signal from the microsphere perfusion of functional vasculature. Scale bars, 10 µm. Six independent experiments with similar results were performed. (C) Self-renewal capacity, a defining characteristic of stem cells, was evaluated by inoculating mice with syngeneic B16 melanomas (2 million cells per mice) together with 15 CFUs of GFP-tagged isolated CD31+CD105+ ECs. After 2 wk of tumor growth, repeated isolations and serial transplantations of lineage depleted single cell suspensions containing the GFP+ tagged ECs and the B16 cells were performed. The figure shows GFP+ blood vessels in the quaternary transplant. ECs were stained for VEGFR-2 (red), vWF (white), and CD31 and CD105 (red). Scale bar, 10 µm. A 3-D reconstitution of a GFP+ blood vessel in the quaternary transplant is also shown (right; a 34-µm thick stack of 34 x–y slices from a confocal scan). Six independent experiments with similar results were performed.

Techniques Used: Expressing, Functional Assay, In Vivo, Flow Cytometry, FACS, Injection, Mouse Assay, Inverted Microscopy, Generated, Transplantation Assay, Staining, Isolation

A genetic deficit in endothelial c-kit expression decreases colony-forming VESCs and results in impaired EC proliferation and angiogenesis, and retardation of tumor growth in vivo. (A) Equivalent lin−CD31+CD105+ EC populations were detected in mutant C57BL/6J mice with a genetic c-kit expression deficit (C57BL/6J-Kit W-sh mice) and in the wt C57BL/6J controls. However, the Kit W-sh mutant mice have very low numbers of CD117+ ECs (here 1% of total lin−CD31+CD105+ ECs). Typical results from FACS analysis of lung ECs are shown. Histograms indicate the percentage of CD117+ cells, control IgG labeling and gatings are also shown. (B) lin−CD31+CD105+ ECs from kit deficient Kit W-sh mutant mice contain abnormally low levels of endothelial CFCs in comparison to wt mice ( p
Figure Legend Snippet: A genetic deficit in endothelial c-kit expression decreases colony-forming VESCs and results in impaired EC proliferation and angiogenesis, and retardation of tumor growth in vivo. (A) Equivalent lin−CD31+CD105+ EC populations were detected in mutant C57BL/6J mice with a genetic c-kit expression deficit (C57BL/6J-Kit W-sh mice) and in the wt C57BL/6J controls. However, the Kit W-sh mutant mice have very low numbers of CD117+ ECs (here 1% of total lin−CD31+CD105+ ECs). Typical results from FACS analysis of lung ECs are shown. Histograms indicate the percentage of CD117+ cells, control IgG labeling and gatings are also shown. (B) lin−CD31+CD105+ ECs from kit deficient Kit W-sh mutant mice contain abnormally low levels of endothelial CFCs in comparison to wt mice ( p

Techniques Used: Expressing, In Vivo, Mutagenesis, Mouse Assay, FACS, Labeling

6) Product Images from "Optimal therapeutic activity of monoclonal antibodies against chikungunya virus requires Fc-FcγR interaction on monocytes"

Article Title: Optimal therapeutic activity of monoclonal antibodies against chikungunya virus requires Fc-FcγR interaction on monocytes

Journal: Science immunology

doi: 10.1126/sciimmunol.aav5062

Clinical protection following mAb therapy. ( A ) Four-week-old WT C57BL/6J mice were administered mouse anti-CHIKV mAbs [CHK-152 + CHK-166 (250 μg per mAb; 500 μg total)] or an isotype control (WNV E60; 500 μg) on 3 dpi with 10 3 FFU of CHIKV. Foot swelling was measured prior to infection and for 10 dpi (n = 8/group, two experiments). Graphs show means ± SEM (***, P
Figure Legend Snippet: Clinical protection following mAb therapy. ( A ) Four-week-old WT C57BL/6J mice were administered mouse anti-CHIKV mAbs [CHK-152 + CHK-166 (250 μg per mAb; 500 μg total)] or an isotype control (WNV E60; 500 μg) on 3 dpi with 10 3 FFU of CHIKV. Foot swelling was measured prior to infection and for 10 dpi (n = 8/group, two experiments). Graphs show means ± SEM (***, P

Techniques Used: Mouse Assay, Infection

7) Product Images from "The obesity-induced adipokine sST2 exacerbates adipose Treg and ILC2 depletion and promotes insulin resistance"

Article Title: The obesity-induced adipokine sST2 exacerbates adipose Treg and ILC2 depletion and promotes insulin resistance

Journal: Science Advances

doi: 10.1126/sciadv.aay6191

Obesity-associated reduction of adipose T regs is linked to elevated expression and secretion of sST2. ( A ) Correlation between eWAT Foxp3 and IL-33 mRNA levels and body weight in a cohort of C57BL/6J mice fed HFD for 8 weeks. ( B ) Schematic diagrams of the ST2 isoforms. Arrows indicate the location of isoform-specific qPCR primers. ( C ) qPCR analysis of sST2 and ST2L expression in a panel of tissues from mice fed chow (filled, n = 4) and HFD (open, n = 4). Data in (C) represent means ± SEM. * P
Figure Legend Snippet: Obesity-associated reduction of adipose T regs is linked to elevated expression and secretion of sST2. ( A ) Correlation between eWAT Foxp3 and IL-33 mRNA levels and body weight in a cohort of C57BL/6J mice fed HFD for 8 weeks. ( B ) Schematic diagrams of the ST2 isoforms. Arrows indicate the location of isoform-specific qPCR primers. ( C ) qPCR analysis of sST2 and ST2L expression in a panel of tissues from mice fed chow (filled, n = 4) and HFD (open, n = 4). Data in (C) represent means ± SEM. * P

Techniques Used: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction

8) Product Images from "A High Serum Iron Level Causes Mouse Retinal Iron Accumulation Despite an Intact Blood-Retinal Barrier"

Article Title: A High Serum Iron Level Causes Mouse Retinal Iron Accumulation Despite an Intact Blood-Retinal Barrier

Journal: The American Journal of Pathology

doi: 10.1016/j.ajpath.2014.07.008

Quantitative PCR results in retina and RPE of i.v. iron sucrose–injected C57BL/6J mice. Relative mRNA levels of Hamp, Tfrc, and L ferritin (Ftl) in the liver ( A–C ), NR ( D–F ), and RPE ( G–I ) of mice with or without iron sucrose
Figure Legend Snippet: Quantitative PCR results in retina and RPE of i.v. iron sucrose–injected C57BL/6J mice. Relative mRNA levels of Hamp, Tfrc, and L ferritin (Ftl) in the liver ( A–C ), NR ( D–F ), and RPE ( G–I ) of mice with or without iron sucrose

Techniques Used: Real-time Polymerase Chain Reaction, Injection, Mouse Assay

Iron labeling in RPE of i.v. iron sucrose–injected C57BL/6J mice. Iron level as determined by immunolabeling for L ferritin (Ftl; red) was greater in the RPE of iron sucrose (V)–injected mice ( B ) compared with controls ( A ). C and D show
Figure Legend Snippet: Iron labeling in RPE of i.v. iron sucrose–injected C57BL/6J mice. Iron level as determined by immunolabeling for L ferritin (Ftl; red) was greater in the RPE of iron sucrose (V)–injected mice ( B ) compared with controls ( A ). C and D show

Techniques Used: Labeling, Injection, Mouse Assay, Immunolabeling

9) Product Images from "Removal of chylomicron remnants in transgenic mice overexpressing normal and membrane-anchored hepatic lipase"

Article Title: Removal of chylomicron remnants in transgenic mice overexpressing normal and membrane-anchored hepatic lipase

Journal: Journal of lipid research

doi: 10.1194/jlr.M400184-JLR200

Plasma lipid concentrations in HL −/− , mHL −/− /rHL +/+ , and mHL −/− /gpi-rHL +/+ mice. Mice were fasted overnight before the blood draw. A: Triglycerides (TG), cholesterol (TC), and phospholipids (PL) were measured as described in Materials and Methods. B: Apolipoprotein B (apoB) and apoA-I were measured by SDS-PAGE as described in Materials and Methods. WT FVB mice were used as controls for both native HL ( mHL −/− /rHL +/+ ) and GPI HL transgenic (tg) ( mHL −/− /gpi-rHL +/+ ) mice. C57BL/6J mice were used as controls for HL knockout (HLKO) ( HL −/− ) mice. * P
Figure Legend Snippet: Plasma lipid concentrations in HL −/− , mHL −/− /rHL +/+ , and mHL −/− /gpi-rHL +/+ mice. Mice were fasted overnight before the blood draw. A: Triglycerides (TG), cholesterol (TC), and phospholipids (PL) were measured as described in Materials and Methods. B: Apolipoprotein B (apoB) and apoA-I were measured by SDS-PAGE as described in Materials and Methods. WT FVB mice were used as controls for both native HL ( mHL −/− /rHL +/+ ) and GPI HL transgenic (tg) ( mHL −/− /gpi-rHL +/+ ) mice. C57BL/6J mice were used as controls for HL knockout (HLKO) ( HL −/− ) mice. * P

Techniques Used: Mouse Assay, SDS Page, Transgenic Assay, Knock-Out

Removal of chylomicron remnants by rat HL transgenic, knockout, and WT mouse livers. 125 I-labeled chylomicron remnants (4 μg/ml perfusate) were added to the perfusate of the isolated livers, and aliquots of the perfusate leaving the liver were collected at 1 min intervals for 20 min. The amount of remnants removed by the liver from the perfusate per pass is expressed as the percentage of 125 I-labeled remnants removed per pass, which is calculated by subtracting the radioactivity in a sample of perfusate that left the liver from the initial radioactivity in the perfusate, divided by the initial radioactivity in the perfusate, and multiplying by 100. A: FVB mice (n = 4), native rat HL transgenic ( mHL −/− /rHL +/+ ) mice (n = 3), native rat HL transgenic mice in the presence of receptor-associated protein (RAP; n = 5). B: FVB mice (n = 4), GPI-anchored rat HL transgenic ( mHL −/− /gpi-rHL +/+ ) mice (n = 3), GPI-anchored rat HL transgenic mice in the presence of RAP (n = 4). C: C57BL/6J mice (n = 4), HL knockout ( HL −/− ) mice (n = 4), HL knockout mice in the presence of RAP (n = 4). * P
Figure Legend Snippet: Removal of chylomicron remnants by rat HL transgenic, knockout, and WT mouse livers. 125 I-labeled chylomicron remnants (4 μg/ml perfusate) were added to the perfusate of the isolated livers, and aliquots of the perfusate leaving the liver were collected at 1 min intervals for 20 min. The amount of remnants removed by the liver from the perfusate per pass is expressed as the percentage of 125 I-labeled remnants removed per pass, which is calculated by subtracting the radioactivity in a sample of perfusate that left the liver from the initial radioactivity in the perfusate, divided by the initial radioactivity in the perfusate, and multiplying by 100. A: FVB mice (n = 4), native rat HL transgenic ( mHL −/− /rHL +/+ ) mice (n = 3), native rat HL transgenic mice in the presence of receptor-associated protein (RAP; n = 5). B: FVB mice (n = 4), GPI-anchored rat HL transgenic ( mHL −/− /gpi-rHL +/+ ) mice (n = 3), GPI-anchored rat HL transgenic mice in the presence of RAP (n = 4). C: C57BL/6J mice (n = 4), HL knockout ( HL −/− ) mice (n = 4), HL knockout mice in the presence of RAP (n = 4). * P

Techniques Used: Transgenic Assay, Knock-Out, Labeling, Isolation, Radioactivity, Mouse Assay

10) Product Images from "IlvY is an important regulator of Shigella infection in vitro and in vivo"

Article Title: IlvY is an important regulator of Shigella infection in vitro and in vivo

Journal: bioRxiv

doi: 10.1101/2020.07.28.220327

IlvY is important for S. sonnei in vivo survival. Wild-type C57Bl/6J mice were infected by oral gavage with 5 x 10 5 CFU S. sonnei that were grown to log-phase in broth culture. At multiple time points post infection, gastrointestinal tissue was collected to characterize the infection burden (A). The observed infection in the large intestine was highest at 2 hours post infection, and all mice remained infected at 24 hours post infection. The inflammatory marker myeloperoxidase was significantly increased in the large intestine by 24 hours and remained elevated at 72 hours post infection (B). Zonulin, a marker of gastrointestinal epithelial permeability, was reduced over the first two days of infection but returned to baseline by 72 hours (C). To confirm whether IlvY has an important role in S. sonnei in vivo survival, Wild-type C57Bl/6J mice were infected by oral gavage with 5 x 10 8 CFU wild-type S. sonnei or Δ ilvY S. sonnei that were grown to log-phase in broth culture. At 24 hours post infection, gastrointestinal tissue was collected to characterize the infection burden (D). Interestingly, mice infected with the Δ ilvY S. sonnei had a significantly lower infection burden compared to mice infected with wild-type Shigella . While the levels of myeloperoxidase were the same in both groups (E), zonulin levels were significantly higher in the large intestine of mice infected with Δ ilvY S. sonnei suggesting that intestinal permeability was not disrupted by infection with the IlvY deficient strain (F).
Figure Legend Snippet: IlvY is important for S. sonnei in vivo survival. Wild-type C57Bl/6J mice were infected by oral gavage with 5 x 10 5 CFU S. sonnei that were grown to log-phase in broth culture. At multiple time points post infection, gastrointestinal tissue was collected to characterize the infection burden (A). The observed infection in the large intestine was highest at 2 hours post infection, and all mice remained infected at 24 hours post infection. The inflammatory marker myeloperoxidase was significantly increased in the large intestine by 24 hours and remained elevated at 72 hours post infection (B). Zonulin, a marker of gastrointestinal epithelial permeability, was reduced over the first two days of infection but returned to baseline by 72 hours (C). To confirm whether IlvY has an important role in S. sonnei in vivo survival, Wild-type C57Bl/6J mice were infected by oral gavage with 5 x 10 8 CFU wild-type S. sonnei or Δ ilvY S. sonnei that were grown to log-phase in broth culture. At 24 hours post infection, gastrointestinal tissue was collected to characterize the infection burden (D). Interestingly, mice infected with the Δ ilvY S. sonnei had a significantly lower infection burden compared to mice infected with wild-type Shigella . While the levels of myeloperoxidase were the same in both groups (E), zonulin levels were significantly higher in the large intestine of mice infected with Δ ilvY S. sonnei suggesting that intestinal permeability was not disrupted by infection with the IlvY deficient strain (F).

Techniques Used: In Vivo, Mouse Assay, Infection, Marker, Permeability

11) Product Images from "Hepatocyte sortilin 1 knockout and treatment with a sortilin 1 inhibitor reduced plasma cholesterol in Western diet-fed mice"

Article Title: Hepatocyte sortilin 1 knockout and treatment with a sortilin 1 inhibitor reduced plasma cholesterol in Western diet-fed mice

Journal: Journal of Lipid Research

doi: 10.1194/jlr.M089789

AF38469 treatment decreased plasma cholesterol and hepatic VLDL secretion in WD-fed mice. A, B, D–F: Male 12-week-old C57BL/6J mice were fed either WD or WD supplemented with AF38469 (AF) for 8 weeks. Plasma cholesterol (A), TG (B), gallbladder (GB) total cholesterol content (D), tissue total bile acid content and bile acid pool (E), and hepatic mRNA expression (E) are shown. C: Male 12-week-old C57BL/6J mice were fed either WD or WD supplemented with AF38469 (AF) for 2 weeks. A VLDL secretion assay was performed as described in the Materials and Methods. Results are expressed as mean ± SEM (n = 5). * P
Figure Legend Snippet: AF38469 treatment decreased plasma cholesterol and hepatic VLDL secretion in WD-fed mice. A, B, D–F: Male 12-week-old C57BL/6J mice were fed either WD or WD supplemented with AF38469 (AF) for 8 weeks. Plasma cholesterol (A), TG (B), gallbladder (GB) total cholesterol content (D), tissue total bile acid content and bile acid pool (E), and hepatic mRNA expression (E) are shown. C: Male 12-week-old C57BL/6J mice were fed either WD or WD supplemented with AF38469 (AF) for 2 weeks. A VLDL secretion assay was performed as described in the Materials and Methods. Results are expressed as mean ± SEM (n = 5). * P

Techniques Used: Mouse Assay, Expressing

AF38469 treatment did not affect fasting glucose or glucose tolerance in WD-fed mice. Male 12-week-old C57BL/6J mice were fed either WD or WD supplemented with AF38469 (AF) for 8 weeks. A: Fasting plasma glucose levels. B: Fasting plasma insulin levels. C: Plasma FFA levels. D: Glucose tolerance test. Results are expressed as mean ± SEM (n = 5). * P
Figure Legend Snippet: AF38469 treatment did not affect fasting glucose or glucose tolerance in WD-fed mice. Male 12-week-old C57BL/6J mice were fed either WD or WD supplemented with AF38469 (AF) for 8 weeks. A: Fasting plasma glucose levels. B: Fasting plasma insulin levels. C: Plasma FFA levels. D: Glucose tolerance test. Results are expressed as mean ± SEM (n = 5). * P

Techniques Used: Mouse Assay

AF38469 treatment did not affect WD-induced weight gain or hepatic steatosis in mice. Male 12-week-old C57BL/6J mice were fed either WD or WD supplemented with AF38469 (AF) for 8 weeks. A: Body weight. B: Liver weight. C: Liver TG content. D. Liver cholesterol content. E: H E staining (scale bar, 100 μm). F, G: Plasma AST and ALT. Results are expressed as mean ± SEM (n = 5).
Figure Legend Snippet: AF38469 treatment did not affect WD-induced weight gain or hepatic steatosis in mice. Male 12-week-old C57BL/6J mice were fed either WD or WD supplemented with AF38469 (AF) for 8 weeks. A: Body weight. B: Liver weight. C: Liver TG content. D. Liver cholesterol content. E: H E staining (scale bar, 100 μm). F, G: Plasma AST and ALT. Results are expressed as mean ± SEM (n = 5).

Techniques Used: Mouse Assay, Staining, AST Assay

12) Product Images from "Proteinase-activated Receptor 2 Activation Promotes an Anti-inflammatory and Alternatively Activated Phenotype in LPS-stimulated Murine Macrophages"

Article Title: Proteinase-activated Receptor 2 Activation Promotes an Anti-inflammatory and Alternatively Activated Phenotype in LPS-stimulated Murine Macrophages

Journal: Innate immunity

doi: 10.1177/1753425910395044

PAR 2 fAP synergistically augments rIL-4- or LPS-induced alternative activation of murine macrophages Thioglycollate-elicited peritoneal macrophage from C57BL/6J mice were stimulated for 48 h with medium (M), PAR 2 fAP (2-furoyl-LIGRLO-NH 2 ; 200 μM), recombinant IL-4 (5 ng ml −1 ), LPS (10 ng ml −1 ), or the indicated treatment combinations. Relative gene expression was analyzed by qPCR. Data are presented as the combined mean ± SEM in two separate experiments. * , p
Figure Legend Snippet: PAR 2 fAP synergistically augments rIL-4- or LPS-induced alternative activation of murine macrophages Thioglycollate-elicited peritoneal macrophage from C57BL/6J mice were stimulated for 48 h with medium (M), PAR 2 fAP (2-furoyl-LIGRLO-NH 2 ; 200 μM), recombinant IL-4 (5 ng ml −1 ), LPS (10 ng ml −1 ), or the indicated treatment combinations. Relative gene expression was analyzed by qPCR. Data are presented as the combined mean ± SEM in two separate experiments. * , p

Techniques Used: Activation Assay, Mouse Assay, Recombinant, Expressing, Real-time Polymerase Chain Reaction

PAR 2 AP differentially modulates LPS-induced cytokine production in primary murine macrophages to promote an anti-inflammatory response Thioglycollate-elicited peritoneal macrophages from C57BL/6J mice were stimulated for 24 h with medium only (Med), PAR 2 AP (SLIGKV-NH 2 ; 200 μM), LPS (100 ng ml −1 ), or LPS in the presence of increasing concentrations of PAR 2 AP (50 – 200 μM). Supernatants were analyzed for cytokine production by ELISA. Data are presented as the mean ± s.d. of duplicate samples and are representative of one of two separate experiments. * , p
Figure Legend Snippet: PAR 2 AP differentially modulates LPS-induced cytokine production in primary murine macrophages to promote an anti-inflammatory response Thioglycollate-elicited peritoneal macrophages from C57BL/6J mice were stimulated for 24 h with medium only (Med), PAR 2 AP (SLIGKV-NH 2 ; 200 μM), LPS (100 ng ml −1 ), or LPS in the presence of increasing concentrations of PAR 2 AP (50 – 200 μM). Supernatants were analyzed for cytokine production by ELISA. Data are presented as the mean ± s.d. of duplicate samples and are representative of one of two separate experiments. * , p

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

PAR 2 AP differentially modulates LPS-induced cytokine gene expression to promote an anti-inflammatory response in vivo C57BL/6J mice were injected i.p. with 0.5 ml of saline, PAR 2 AP (SLIGKV-NH 2 ; 500 μM), LPS (25 μg), or both. At the indicated times, livers were harvested, and relative gene expression was analyzed by qPCR. The results are the mean ± s.e.m (n = 3 for each group at each time point) of a representative experiment (N = 2). *, p
Figure Legend Snippet: PAR 2 AP differentially modulates LPS-induced cytokine gene expression to promote an anti-inflammatory response in vivo C57BL/6J mice were injected i.p. with 0.5 ml of saline, PAR 2 AP (SLIGKV-NH 2 ; 500 μM), LPS (25 μg), or both. At the indicated times, livers were harvested, and relative gene expression was analyzed by qPCR. The results are the mean ± s.e.m (n = 3 for each group at each time point) of a representative experiment (N = 2). *, p

Techniques Used: Expressing, In Vivo, Mouse Assay, Injection, Real-time Polymerase Chain Reaction

13) Product Images from "Complete FcRn dependence for intravenous Ig therapy in autoimmune skin blistering diseases"

Article Title: Complete FcRn dependence for intravenous Ig therapy in autoimmune skin blistering diseases

Journal:

doi: 10.1172/JCI24394

HDIG inhibits experimental PF and PV by accelerating degradation of pathogenic human autoantibody IgG. Neonatal C57BL/6J mice were pretreated with buffer, IgM (12.93 mg/g body weight), or different doses of HDIG (0–2 mg/g body weight) and injected
Figure Legend Snippet: HDIG inhibits experimental PF and PV by accelerating degradation of pathogenic human autoantibody IgG. Neonatal C57BL/6J mice were pretreated with buffer, IgM (12.93 mg/g body weight), or different doses of HDIG (0–2 mg/g body weight) and injected

Techniques Used: Mouse Assay, Injection

High-dose Ig inhibits experimental BP by accelerating degradation of pathogenic anti-mBP180 IgG. Neonatal C57BL/6J mice were pretreated with buffer, IgM (12.93 mg/g body weight), or different doses of HDIG (0–2 mg/g body weight) and injected i.p.
Figure Legend Snippet: High-dose Ig inhibits experimental BP by accelerating degradation of pathogenic anti-mBP180 IgG. Neonatal C57BL/6J mice were pretreated with buffer, IgM (12.93 mg/g body weight), or different doses of HDIG (0–2 mg/g body weight) and injected i.p.

Techniques Used: Mouse Assay, Injection

14) Product Images from "Fever Promotes T Lymphocyte Trafficking via a Thermal Sensory Pathway Involving Heat Shock Protein 90 and α4 Integrins"

Article Title: Fever Promotes T Lymphocyte Trafficking via a Thermal Sensory Pathway Involving Heat Shock Protein 90 and α4 Integrins

Journal: Immunity

doi: 10.1016/j.immuni.2018.11.013

Disruption of Hsp90-α4 Interaction Inhibits Fever-Induced T Cell Trafficking In vivo and Impairs the Clearance of Bacterial Infection (A–D) WT and KI C57BL/6J mice were treated with normothermia (NT; core temperature 36.8°C ± 0.2°C) or fever-range whole-body hyperthermia (WBH; core temperature 39.5°C ± 0.5°C) for 6 hr (n = 7–10 mice per group) and then were sacrificed. T cells were isolated from the spleen. α4β7-VCAM-1 binding was disrupted by cell pre-treatment with 10 μg/mL α4β7-blocking antibody DATK32 during examination of α4β1-mediated cell adhesion and migration on VCAM-1 substrate in (B) and (C). Co-immunoprecipitation of Hsp90AA1 and Hsp90AB1 with integrin α4 in the cell-membrane fractions is shown in (A). Adhesion of T cells to immobilized VCAM-1-Fc (5 μg/mL) or MAdCAM-1-Fc (5 μg/mL) substrate in 1 mM Ca 2+ + Mg 2+ at a wall shear stress of 1 dyn/cm 2 is shown in (B). Transmigration of T cells across membranes coated in VCAM-1-Fc (5 μg/mL) or MAdCAM-1-Fc (5 μg/mL) in the presence of CCL21 (500 ng/mL) in the lower chamber is shown in (C). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified (D). (E–G) WT and KI mice were injected with LPS (10 μg/kg) or PBS at time zero (n = 3 mice per group). Body temperature was monitored every hour. The bar shows the dark period (E). Immunoblot analysis of Hsp90AA1 and Hsp90AB1 in T cells isolated from PLNs in mice is shown in (F). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified (G). (H) Effect of different temperatures on the expression of Hsp90. T cells were isolated from PLNs in WT mice and then treated at 37°C, 38°C, 38.5°C, 39°C, or 40°C for 12 hr. Immunoblot analysis of Hsp90AA1 and Hsp90AB1 in T cells is shown. (I–M) WT and KI mice were orally injected with PBS or S. typhimurium strain SL1344 (10 8 CFU per mouse; n = 4–12 mice per group). Body temperature was monitored for 5 days (I). The survival rates of WT and KI mice are shown in (J), and significance was calculated as given. H E staining of the small intestine at day 5 after infection is shown in (K) (scale bar, 100 mm). Immunofluorescence analysis of the small intestine sections at day 5 after infection is shown. Stain coloring is as follows: DAPI, blue; S. typhimurium expressing GFP, green; and CD3, red. Quantifications of S. typhimurium colonies and CD3 + cells are shown below in (L) (scale bar, 100 mm). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified at day 5 after infection (M). .
Figure Legend Snippet: Disruption of Hsp90-α4 Interaction Inhibits Fever-Induced T Cell Trafficking In vivo and Impairs the Clearance of Bacterial Infection (A–D) WT and KI C57BL/6J mice were treated with normothermia (NT; core temperature 36.8°C ± 0.2°C) or fever-range whole-body hyperthermia (WBH; core temperature 39.5°C ± 0.5°C) for 6 hr (n = 7–10 mice per group) and then were sacrificed. T cells were isolated from the spleen. α4β7-VCAM-1 binding was disrupted by cell pre-treatment with 10 μg/mL α4β7-blocking antibody DATK32 during examination of α4β1-mediated cell adhesion and migration on VCAM-1 substrate in (B) and (C). Co-immunoprecipitation of Hsp90AA1 and Hsp90AB1 with integrin α4 in the cell-membrane fractions is shown in (A). Adhesion of T cells to immobilized VCAM-1-Fc (5 μg/mL) or MAdCAM-1-Fc (5 μg/mL) substrate in 1 mM Ca 2+ + Mg 2+ at a wall shear stress of 1 dyn/cm 2 is shown in (B). Transmigration of T cells across membranes coated in VCAM-1-Fc (5 μg/mL) or MAdCAM-1-Fc (5 μg/mL) in the presence of CCL21 (500 ng/mL) in the lower chamber is shown in (C). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified (D). (E–G) WT and KI mice were injected with LPS (10 μg/kg) or PBS at time zero (n = 3 mice per group). Body temperature was monitored every hour. The bar shows the dark period (E). Immunoblot analysis of Hsp90AA1 and Hsp90AB1 in T cells isolated from PLNs in mice is shown in (F). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified (G). (H) Effect of different temperatures on the expression of Hsp90. T cells were isolated from PLNs in WT mice and then treated at 37°C, 38°C, 38.5°C, 39°C, or 40°C for 12 hr. Immunoblot analysis of Hsp90AA1 and Hsp90AB1 in T cells is shown. (I–M) WT and KI mice were orally injected with PBS or S. typhimurium strain SL1344 (10 8 CFU per mouse; n = 4–12 mice per group). Body temperature was monitored for 5 days (I). The survival rates of WT and KI mice are shown in (J), and significance was calculated as given. H E staining of the small intestine at day 5 after infection is shown in (K) (scale bar, 100 mm). Immunofluorescence analysis of the small intestine sections at day 5 after infection is shown. Stain coloring is as follows: DAPI, blue; S. typhimurium expressing GFP, green; and CD3, red. Quantifications of S. typhimurium colonies and CD3 + cells are shown below in (L) (scale bar, 100 mm). The total numbers of T cells in PLNs, MLNs, PPs, spleen, and PB were quantified at day 5 after infection (M). .

Techniques Used: In Vivo, Infection, Mouse Assay, Isolation, Binding Assay, Blocking Assay, Migration, Immunoprecipitation, Transmigration Assay, Injection, Expressing, Staining, Immunofluorescence

15) Product Images from "IlvY is an important regulator of Shigella infection in vitro and in vivo"

Article Title: IlvY is an important regulator of Shigella infection in vitro and in vivo

Journal: bioRxiv

doi: 10.1101/2020.07.28.220327

IlvY is important for S. sonnei in vivo survival. Wild-type C57Bl/6J mice were infected by oral gavage with 5 x 10 5 CFU S. sonnei that were grown to log-phase in broth culture. At multiple time points post infection, gastrointestinal tissue was collected to characterize the infection burden (A). The observed infection in the large intestine was highest at 2 hours post infection, and all mice remained infected at 24 hours post infection. The inflammatory marker myeloperoxidase was significantly increased in the large intestine by 24 hours and remained elevated at 72 hours post infection (B). Zonulin, a marker of gastrointestinal epithelial permeability, was reduced over the first two days of infection but returned to baseline by 72 hours (C). To confirm whether IlvY has an important role in S. sonnei in vivo survival, Wild-type C57Bl/6J mice were infected by oral gavage with 5 x 10 8 CFU wild-type S. sonnei or Δ ilvY S. sonnei that were grown to log-phase in broth culture. At 24 hours post infection, gastrointestinal tissue was collected to characterize the infection burden (D). Interestingly, mice infected with the Δ ilvY S. sonnei had a significantly lower infection burden compared to mice infected with wild-type Shigella . While the levels of myeloperoxidase were the same in both groups (E), zonulin levels were significantly higher in the large intestine of mice infected with Δ ilvY S. sonnei suggesting that intestinal permeability was not disrupted by infection with the IlvY deficient strain (F).
Figure Legend Snippet: IlvY is important for S. sonnei in vivo survival. Wild-type C57Bl/6J mice were infected by oral gavage with 5 x 10 5 CFU S. sonnei that were grown to log-phase in broth culture. At multiple time points post infection, gastrointestinal tissue was collected to characterize the infection burden (A). The observed infection in the large intestine was highest at 2 hours post infection, and all mice remained infected at 24 hours post infection. The inflammatory marker myeloperoxidase was significantly increased in the large intestine by 24 hours and remained elevated at 72 hours post infection (B). Zonulin, a marker of gastrointestinal epithelial permeability, was reduced over the first two days of infection but returned to baseline by 72 hours (C). To confirm whether IlvY has an important role in S. sonnei in vivo survival, Wild-type C57Bl/6J mice were infected by oral gavage with 5 x 10 8 CFU wild-type S. sonnei or Δ ilvY S. sonnei that were grown to log-phase in broth culture. At 24 hours post infection, gastrointestinal tissue was collected to characterize the infection burden (D). Interestingly, mice infected with the Δ ilvY S. sonnei had a significantly lower infection burden compared to mice infected with wild-type Shigella . While the levels of myeloperoxidase were the same in both groups (E), zonulin levels were significantly higher in the large intestine of mice infected with Δ ilvY S. sonnei suggesting that intestinal permeability was not disrupted by infection with the IlvY deficient strain (F).

Techniques Used: In Vivo, Mouse Assay, Infection, Marker, Permeability

16) Product Images from "Tauroursodeoxycholic acid preserves photoreceptor structure and function in the rd10 mouse through post-natal day 30"

Article Title: Tauroursodeoxycholic acid preserves photoreceptor structure and function in the rd10 mouse through post-natal day 30

Journal:

doi: 10.1167/iovs.07-1012

Studies in WT mice: Retinal function measurements were recorded from WT C57BL/6J mice treated with TUDCA or vehicle from P6 to P30. A) Representative dark-adapted ERG waveforms for WT TUDCA- and vehicle-treated mice. Dark-adapted a-wave (B), dark-adapted
Figure Legend Snippet: Studies in WT mice: Retinal function measurements were recorded from WT C57BL/6J mice treated with TUDCA or vehicle from P6 to P30. A) Representative dark-adapted ERG waveforms for WT TUDCA- and vehicle-treated mice. Dark-adapted a-wave (B), dark-adapted

Techniques Used: Mouse Assay

17) Product Images from "Antisense suppression of glial fibrillary acidic protein as a treatment for Alexander disease"

Article Title: Antisense suppression of glial fibrillary acidic protein as a treatment for Alexander disease

Journal: Annals of neurology

doi: 10.1002/ana.25118

Suppression of GFAP in AxD mice mitigates molecular, cellular, and clinical phenotypes. ( A ) Weekly body weights for Gfap +/+ and Gfap +/R236H mice treated at 8 weeks of age with saline, ASO-4 or ASO-9 (300 μg ASO, n = 4–6 males). P values represent 2-way RM ANOVA comparison between saline and ASO treated Gfap +/R236H mice. Asterisks indicate significant differences as indicated by Sidak’s multiple comparisons, error bars = standard deviation. ( B ) Western analysis of phosphorylated STAT3-Y705 shows decreased activation in Gfap +/R236H mice treated with 500 μg ASO-5 versus saline at 8 weeks post treatment (hippocampus shown, n = 3–4 C57BL/6J mice, males and females treated at 8 weeks of age, asterisks represent 2-tailed unpaired t-test comparing saline versus ASO-treated Gfap +/R236H mice, ****p
Figure Legend Snippet: Suppression of GFAP in AxD mice mitigates molecular, cellular, and clinical phenotypes. ( A ) Weekly body weights for Gfap +/+ and Gfap +/R236H mice treated at 8 weeks of age with saline, ASO-4 or ASO-9 (300 μg ASO, n = 4–6 males). P values represent 2-way RM ANOVA comparison between saline and ASO treated Gfap +/R236H mice. Asterisks indicate significant differences as indicated by Sidak’s multiple comparisons, error bars = standard deviation. ( B ) Western analysis of phosphorylated STAT3-Y705 shows decreased activation in Gfap +/R236H mice treated with 500 μg ASO-5 versus saline at 8 weeks post treatment (hippocampus shown, n = 3–4 C57BL/6J mice, males and females treated at 8 weeks of age, asterisks represent 2-tailed unpaired t-test comparing saline versus ASO-treated Gfap +/R236H mice, ****p

Techniques Used: Mouse Assay, Allele-specific Oligonucleotide, Standard Deviation, Western Blot, Activation Assay

18) Product Images from "Novel Strategies for Targeting Innate Immune Responses to Influenza"

Article Title: Novel Strategies for Targeting Innate Immune Responses to Influenza

Journal: Mucosal immunology

doi: 10.1038/mi.2015.141

Effect of IL-1Ra against lethal influenza challenge. C57BL/6J mice were infected with mouse-adapted influenza strain PR8 (∼7500 TCID 50 , i.n.). Mice received vehicle (saline; i.v.), Eritoran (E5564; 200 μg/mouse; i.v.), or the IL-1Ra (150 μg/mouse; i.v.) from day 2 to day 6 post-infection. Survival (top panel) and clinical scores (bottom panel) were monitored daily for 14 days. Data shown is combined from 2 separate experiments (5 mice/treatment group/experiment).
Figure Legend Snippet: Effect of IL-1Ra against lethal influenza challenge. C57BL/6J mice were infected with mouse-adapted influenza strain PR8 (∼7500 TCID 50 , i.n.). Mice received vehicle (saline; i.v.), Eritoran (E5564; 200 μg/mouse; i.v.), or the IL-1Ra (150 μg/mouse; i.v.) from day 2 to day 6 post-infection. Survival (top panel) and clinical scores (bottom panel) were monitored daily for 14 days. Data shown is combined from 2 separate experiments (5 mice/treatment group/experiment).

Techniques Used: Mouse Assay, Infection

Anti-TLR2 IgG treatment protects mice from lethal influenza challenge. (A) Experimental protocol. C57BL/6J mice were either treated with isotype control IgG or anti-TLR2 (T2.5; 100 μg/ms; i.v.) 3 h prior to and 1 day post-infection or on days 2 and 4 post-infection. Survival (B) was monitored daily. Data shown is combined results of 2 separate experiments (5 mice/treatment group/experiment).
Figure Legend Snippet: Anti-TLR2 IgG treatment protects mice from lethal influenza challenge. (A) Experimental protocol. C57BL/6J mice were either treated with isotype control IgG or anti-TLR2 (T2.5; 100 μg/ms; i.v.) 3 h prior to and 1 day post-infection or on days 2 and 4 post-infection. Survival (B) was monitored daily. Data shown is combined results of 2 separate experiments (5 mice/treatment group/experiment).

Techniques Used: Mouse Assay, Mass Spectrometry, Infection

Effect of AT-1001 against lethal influenza challenge. (A) C57BL/6J mice were infected with mouse-adapted influenza strain PR8 (∼7500 TCID 50 , i.n.; ∼LD 90 ). Mice received vehicle (saline; i.v.), Eritoran (E5564; 200 μg/mouse; i.v.), or AT-1001 (150 μg/mouse; i.v.) from day 2 to day 6 post-infection. Survival (A) and clinical scores (B) were monitored daily for 14 days. Data shown is combined from 3 separate experiments (5 mice/treatment group/experiment). (C) Lung wet-to-dry (W/D) weight ratio as an index for pulmonary edema after infection. C57BL/6J mice were infected and treated as described above. On day 7 post-infection, lungs were harvested and and lung weights were measured immediately after excision and recorded as wet weight. Lung tissue was air dried for 5-6 days and re-weighed until a stable dry weight obtained. The W/D weight ratio was calculated by dividing the wet by dry weight. The N for each group is indicated in each bar. Each vertical bar represents the mean ( + s.e.m.).
Figure Legend Snippet: Effect of AT-1001 against lethal influenza challenge. (A) C57BL/6J mice were infected with mouse-adapted influenza strain PR8 (∼7500 TCID 50 , i.n.; ∼LD 90 ). Mice received vehicle (saline; i.v.), Eritoran (E5564; 200 μg/mouse; i.v.), or AT-1001 (150 μg/mouse; i.v.) from day 2 to day 6 post-infection. Survival (A) and clinical scores (B) were monitored daily for 14 days. Data shown is combined from 3 separate experiments (5 mice/treatment group/experiment). (C) Lung wet-to-dry (W/D) weight ratio as an index for pulmonary edema after infection. C57BL/6J mice were infected and treated as described above. On day 7 post-infection, lungs were harvested and and lung weights were measured immediately after excision and recorded as wet weight. Lung tissue was air dried for 5-6 days and re-weighed until a stable dry weight obtained. The W/D weight ratio was calculated by dividing the wet by dry weight. The N for each group is indicated in each bar. Each vertical bar represents the mean ( + s.e.m.).

Techniques Used: Mouse Assay, Infection

Anti-TLR4 IgG treatment protects mice from lethal influenza challenge. (A) C57BL/6J mice were infected with mouse-adapted influenza strain PR8 (∼7500 TCID 50 , i.n.). Mice received either control IgG or a highly specific anti-TLR4 IgG (2 mg/mouse; i.v.) once (day 2 only) or twice (days 2 and 4). Survival (B) and clinical scores (C) were monitored daily. Each graph represents the combined results of 2 separate experiments (5 mice/treatment group/experiment).
Figure Legend Snippet: Anti-TLR4 IgG treatment protects mice from lethal influenza challenge. (A) C57BL/6J mice were infected with mouse-adapted influenza strain PR8 (∼7500 TCID 50 , i.n.). Mice received either control IgG or a highly specific anti-TLR4 IgG (2 mg/mouse; i.v.) once (day 2 only) or twice (days 2 and 4). Survival (B) and clinical scores (C) were monitored daily. Each graph represents the combined results of 2 separate experiments (5 mice/treatment group/experiment).

Techniques Used: Mouse Assay, Infection

19) Product Images from "Novel Role of Gastric Releasing Peptide (GRP)-Mediated Signaling in the Host Response to Influenza Infection"

Article Title: Novel Role of Gastric Releasing Peptide (GRP)-Mediated Signaling in the Host Response to Influenza Infection

Journal: Mucosal immunology

doi: 10.1038/s41385-018-0081-9

Blocking GRP blunts influenza-induced cytokine induction in vivo . WT C57BL/6J mice (5 mice/treatment group) were infected i.n. with mouse-adapted influenza strain PR8 (LD 90 ; ~7500 TCID 50 ). Mice received vehicle (saline+0.0096% DMSO) or GRP inhibitor (NSC77427; 20 μM/mouse; i.v.) daily from day 2 to day 6 post-infection. On day 7 post-infection, lungs were harvested and total RNA was extracted to measure gene expression by qRT-PCR. ** p
Figure Legend Snippet: Blocking GRP blunts influenza-induced cytokine induction in vivo . WT C57BL/6J mice (5 mice/treatment group) were infected i.n. with mouse-adapted influenza strain PR8 (LD 90 ; ~7500 TCID 50 ). Mice received vehicle (saline+0.0096% DMSO) or GRP inhibitor (NSC77427; 20 μM/mouse; i.v.) daily from day 2 to day 6 post-infection. On day 7 post-infection, lungs were harvested and total RNA was extracted to measure gene expression by qRT-PCR. ** p

Techniques Used: Blocking Assay, In Vivo, Mouse Assay, Infection, Expressing, Quantitative RT-PCR

Influenza infection of mice and cotton rats induces GRP in lungs and serum. (A) WT C57BL/6J mice were infected with mouse-adapted influenza strain PR8 (LD 90 ; ~7500 TCID 50 ). Mice were euthanized on days 0, 2, 4, 6, and 8 post-infection (3–5 mice/group; * p
Figure Legend Snippet: Influenza infection of mice and cotton rats induces GRP in lungs and serum. (A) WT C57BL/6J mice were infected with mouse-adapted influenza strain PR8 (LD 90 ; ~7500 TCID 50 ). Mice were euthanized on days 0, 2, 4, 6, and 8 post-infection (3–5 mice/group; * p

Techniques Used: Infection, Mouse Assay

Blocking GRP and GRPR enhances survival after influenza infection. (A) WT C57BL/6J mice were infected with mouse-adapted influenza strain PR8 (LD 90 ; ~7500 TCID 50 ). Mice received vehicle (saline + 0.096% DMSO) or GRP inhibitor (NSC77427; 20 μM, 100 μl i.v./mouse) daily from day 2 to day 6 postinfection. Survival was monitored for 14 days. Data shown are combined results of 3 assays (5 mice/treatment group/experiment). (B) WT C57BL/6J mice were infected as described in (A). Mice received either control IgG or a highly specific anti-GRP IgG (100 μg; 100 μl i.v./mouse) on day 2 and day 4 post-PR8 infection. Survival was monitored as in (A). Data shown are combined results of 2 assays (5 mice/treatment group/experiment). (C) WT C57BL/6J mice were infected as described in (A). Mice received vehicle (saline) or GRPR antagonist (BW2258U89; 20 μM, 100 μl i.v./mouse). Survival was monitored as in (A). Data shown are combined results of 2 assays (5 mice/treatment group/experiment).
Figure Legend Snippet: Blocking GRP and GRPR enhances survival after influenza infection. (A) WT C57BL/6J mice were infected with mouse-adapted influenza strain PR8 (LD 90 ; ~7500 TCID 50 ). Mice received vehicle (saline + 0.096% DMSO) or GRP inhibitor (NSC77427; 20 μM, 100 μl i.v./mouse) daily from day 2 to day 6 postinfection. Survival was monitored for 14 days. Data shown are combined results of 3 assays (5 mice/treatment group/experiment). (B) WT C57BL/6J mice were infected as described in (A). Mice received either control IgG or a highly specific anti-GRP IgG (100 μg; 100 μl i.v./mouse) on day 2 and day 4 post-PR8 infection. Survival was monitored as in (A). Data shown are combined results of 2 assays (5 mice/treatment group/experiment). (C) WT C57BL/6J mice were infected as described in (A). Mice received vehicle (saline) or GRPR antagonist (BW2258U89; 20 μM, 100 μl i.v./mouse). Survival was monitored as in (A). Data shown are combined results of 2 assays (5 mice/treatment group/experiment).

Techniques Used: Blocking Assay, Infection, Mouse Assay

GRP inhibitor, NSC77427, reduces lung pathology after influenza PR8 challenge. WT C57BL/6J mice (5 mice/treatment group) were infected i.n. with mouse-adapted influenza strain PR8 (LD 90 ; ~7500 TCID 50 ). Mice received vehicle (saline+0.0096% DMSO) or GRP inhibitor (NSC77427; 20 μM/mouse; i.v.) daily from day 2 to day 6 post-infection. On day 7 post-infection, mice were euthanized and lungs were extracted, fixed, and stained for histopathology. Photomicrographs of representative sections were taken at 10x (A, B, and D), and at 40x (C). All scale bars are 100 microns. N = 4 mice/group. * indicates the pleural surface. (E) Quantitation of lung injury is based on the scoring system detailed in the Materials and Methods section. Data shown are mean ± SD. *, p = 0.002
Figure Legend Snippet: GRP inhibitor, NSC77427, reduces lung pathology after influenza PR8 challenge. WT C57BL/6J mice (5 mice/treatment group) were infected i.n. with mouse-adapted influenza strain PR8 (LD 90 ; ~7500 TCID 50 ). Mice received vehicle (saline+0.0096% DMSO) or GRP inhibitor (NSC77427; 20 μM/mouse; i.v.) daily from day 2 to day 6 post-infection. On day 7 post-infection, mice were euthanized and lungs were extracted, fixed, and stained for histopathology. Photomicrographs of representative sections were taken at 10x (A, B, and D), and at 40x (C). All scale bars are 100 microns. N = 4 mice/group. * indicates the pleural surface. (E) Quantitation of lung injury is based on the scoring system detailed in the Materials and Methods section. Data shown are mean ± SD. *, p = 0.002

Techniques Used: Mouse Assay, Infection, Staining, Histopathology, Quantitation Assay

20) Product Images from "Chikungunya Virus Evades Antiviral CD8+ T Cell Responses To Establish Persistent Infection in Joint-Associated Tissues"

Article Title: Chikungunya Virus Evades Antiviral CD8+ T Cell Responses To Establish Persistent Infection in Joint-Associated Tissues

Journal: Journal of Virology

doi: 10.1128/JVI.02036-19

Antigen-specific CD8 + T cells induced in CHIKV(OVA)-AF infection express low levels of the inhibitory receptors PD-1, Tim-3, and Lag-3. (A and C) WT C57BL/6J mice ( n = 4 mice/group) were inoculated i.p. with 2 × 10 5 PFU LCMV-Arm, i.v. with 2 × 10 6 PFU LCMV-cl13, or s.c. in the left foot with 10 3 PFU CHIKV(OVA)-AF. At 10 dpi, PD-1, Tim-3, and Lag-3 expression on antigen-specific CD8 + T cells in the spleen was quantified by flow cytometry. (A) Representative histograms showing PD-1, Tim-3, and Lag-3 expression. PD-1-, Tim-3-, and Lag-3-negative cells are represented by CD45 + B220 − TCRβ + CD8α + CD44 lo tet − cells from LCMV-Arm-infected mice. The remaining histograms are gated on antigen-specific CD8 + T cells (CD45 + B220 − TCRβ + CD8α + CD44 hi tet + ), D b GP 33 + cells (LCMV-Arm- and LCMV-cl13-infected mice), or K b OVA 257 + cells [CHIKV(OVA)-AF-infected mice]. The numbers embedded in the histograms represent the average geometric mean fluorescence intensity (GMFI) among tetramer-positive cells and are summarized in panel C. (B and D) WT C57BL/6J mice ( n = 3 to 4 mice/group) were inoculated s.c. in the left foot with 10 3 PFU LCMV-Arm or CHIKV(OVA)-AF. At 10 dpi, PD-1, Tim-3, and Lag-3 expression on antigen-specific CD8 + T cells in the ipsilateral (I.) ankle tissue was assessed by flow cytometry. (B) Representative histograms showing PD-1, Tim-3, and Lag-3 expression on antigen-specific CD8 + T cells. Histograms are gated on antigen-specific CD8 + T cells (CD45 + B220 − TCRβ + CD8α + CD44 hi Tet + ), D b GP 33 + cells (LCMV-Arm-infected mice), or K b OVA 257 + cells [CHIKV(OVA)-AF-infected mice]. The numbers embedded in the histograms represent the average geometric mean fluorescence intensity among tetramer-positive cells and are summarized in panel D. The data in the graphs represent the mean ± SEM. Statistically significant differences were determined by one-way analysis of variance with Tukey’s multiple-comparison test (C) or by unpaired Student's t test (D). *, P
Figure Legend Snippet: Antigen-specific CD8 + T cells induced in CHIKV(OVA)-AF infection express low levels of the inhibitory receptors PD-1, Tim-3, and Lag-3. (A and C) WT C57BL/6J mice ( n = 4 mice/group) were inoculated i.p. with 2 × 10 5 PFU LCMV-Arm, i.v. with 2 × 10 6 PFU LCMV-cl13, or s.c. in the left foot with 10 3 PFU CHIKV(OVA)-AF. At 10 dpi, PD-1, Tim-3, and Lag-3 expression on antigen-specific CD8 + T cells in the spleen was quantified by flow cytometry. (A) Representative histograms showing PD-1, Tim-3, and Lag-3 expression. PD-1-, Tim-3-, and Lag-3-negative cells are represented by CD45 + B220 − TCRβ + CD8α + CD44 lo tet − cells from LCMV-Arm-infected mice. The remaining histograms are gated on antigen-specific CD8 + T cells (CD45 + B220 − TCRβ + CD8α + CD44 hi tet + ), D b GP 33 + cells (LCMV-Arm- and LCMV-cl13-infected mice), or K b OVA 257 + cells [CHIKV(OVA)-AF-infected mice]. The numbers embedded in the histograms represent the average geometric mean fluorescence intensity (GMFI) among tetramer-positive cells and are summarized in panel C. (B and D) WT C57BL/6J mice ( n = 3 to 4 mice/group) were inoculated s.c. in the left foot with 10 3 PFU LCMV-Arm or CHIKV(OVA)-AF. At 10 dpi, PD-1, Tim-3, and Lag-3 expression on antigen-specific CD8 + T cells in the ipsilateral (I.) ankle tissue was assessed by flow cytometry. (B) Representative histograms showing PD-1, Tim-3, and Lag-3 expression on antigen-specific CD8 + T cells. Histograms are gated on antigen-specific CD8 + T cells (CD45 + B220 − TCRβ + CD8α + CD44 hi Tet + ), D b GP 33 + cells (LCMV-Arm-infected mice), or K b OVA 257 + cells [CHIKV(OVA)-AF-infected mice]. The numbers embedded in the histograms represent the average geometric mean fluorescence intensity among tetramer-positive cells and are summarized in panel D. The data in the graphs represent the mean ± SEM. Statistically significant differences were determined by one-way analysis of variance with Tukey’s multiple-comparison test (C) or by unpaired Student's t test (D). *, P

Techniques Used: Infection, Mouse Assay, Expressing, Flow Cytometry, Fluorescence

The absence of CD8 + T cells impairs the clearance of LCMV in joint-associated and lymphoid tissue. (A and B) WT C57BL/6J mice were inoculated with 2 × 10 5 PFU LCMV-Arm via the i.p. route ( n = 5 mice) or 2 × 10 6 PFU LCMV-cl13 via the i.v. route ( n = 5 mice). (A) Weight gain was monitored daily and is reported as a percentage of the starting weight. (B) At 10 dpi, viral loads in the serum, spleen, liver, and kidney were quantified by plaque assay. Bars represent the SEM. Statistically significant differences were determined by two-way analysis of variance with Bonferroni’s posttest (A) or by Mann-Whitney test (B). **, P
Figure Legend Snippet: The absence of CD8 + T cells impairs the clearance of LCMV in joint-associated and lymphoid tissue. (A and B) WT C57BL/6J mice were inoculated with 2 × 10 5 PFU LCMV-Arm via the i.p. route ( n = 5 mice) or 2 × 10 6 PFU LCMV-cl13 via the i.v. route ( n = 5 mice). (A) Weight gain was monitored daily and is reported as a percentage of the starting weight. (B) At 10 dpi, viral loads in the serum, spleen, liver, and kidney were quantified by plaque assay. Bars represent the SEM. Statistically significant differences were determined by two-way analysis of variance with Bonferroni’s posttest (A) or by Mann-Whitney test (B). **, P

Techniques Used: Mouse Assay, Plaque Assay, MANN-WHITNEY

Antigen-specific CD8 + T cells induced during CHIKV(OVA)-AF infection have polyfunctional effector responses. WT C57BL/6J mice ( n = 4 to 12 mice/group) were inoculated in the left foot with PBS (mock) or 10 3 PFU of CHIKV(OVA)-AF. Spleen cells (5 or 10 dpi) were stimulated with the OVA 257 or E1 331 peptide in the presence of anti-CD107a and brefeldin A for 5 h at 37°C. (A) Representative flow cytometry plots showing IFN-γ production in CD8 + T cells following OVA 257 or E1 331 peptide stimulation. The plots are gated on CD8 + T cells (CD45 + B220 − TCRβ + CD8α + ). The numbers embedded in the plots represent the frequency of IFN-γ + cells among gated CD8 + T cells. (B) Frequency of CD8 + T cells (CD45 + B220 − TCRβ + CD8α + ) producing IFN-γ. (C) Total numbers of OVA 257 - or E1 331 -specific IFN-γ + CD8 + T cells (CD45 + B220 − TCRβ + CD8α + IFN-γ + ). (D) Representative dot plots from 10 dpi gated on CD45 + B220 − TCRβ + CD8α + T cells. The numbers embedded in the plots represent the frequency among all gated cells. Polyfunctional cytokine-producing responder CD8 + T cells are classified as IFN-γ single positive (IFN-γ + ), TNF-α single positive (TNF-α + ), or IFN-γ and TNF-α double positive (IFN-γ + TNF-α + ). The pie charts indicate the percentage of each responding population among all responders. (E) Polyfunctional analysis showing the cytokine production and the cytolytic potential of responding CD8 + T cells. Dot plots from 10 dpi are gated on CD45 + B220 − TCRβ + CD8α + T cells, and the numbers embedded in the plots represent the frequency among all gated cells. Responders were classified as IFN-γ + single positive (IFN-γ + ), CD107a single positive (CD107a + ), or IFN-γ + cells that were degranulated (IFN-γ + CD107a + ) in response to OVA 257 or E1 331 peptide stimulation. The pie charts indicate the percentage of each responding population among all responders. (F to H) At 10 dpi, leukocytes were isolated from ipsilateral and contralateral ankle tissue and stimulated with OVA 257 or E1 331 peptide in the presence of brefeldin A for 5 h at 37°C. Mock samples represent cells combined from ipsilateral and contralateral tissues. (F) Representative flow plots from ipsilateral ankle tissue showing IFN-γ production by antigen-specific CD8 + T cells in response to OVA 257 or E1 331 peptide stimulation. The numbers embedded in the flow plots represent the frequency of CD8 + T cells (CD45 + B220 − TCRβ + CD8α + ) producing IFN-γ. (G) Frequency of cells producing IFN-γ among all CD8 + T cells. (H) Total OVA 257 - and E1 331 -specific IFN-γ + CD8 + T cells. The data in the graphs represent the mean ± SEM. Data were combined from 2 to 3 independent experiments. Statistically significant differences between mock- and CHIKV(OVA)-AF-infected mice were determined by one-way analysis of variance with Tukey’s posttest (B and C) or by the Kruskal-Wallis test with Dunn’s posttest (G and H). *, P
Figure Legend Snippet: Antigen-specific CD8 + T cells induced during CHIKV(OVA)-AF infection have polyfunctional effector responses. WT C57BL/6J mice ( n = 4 to 12 mice/group) were inoculated in the left foot with PBS (mock) or 10 3 PFU of CHIKV(OVA)-AF. Spleen cells (5 or 10 dpi) were stimulated with the OVA 257 or E1 331 peptide in the presence of anti-CD107a and brefeldin A for 5 h at 37°C. (A) Representative flow cytometry plots showing IFN-γ production in CD8 + T cells following OVA 257 or E1 331 peptide stimulation. The plots are gated on CD8 + T cells (CD45 + B220 − TCRβ + CD8α + ). The numbers embedded in the plots represent the frequency of IFN-γ + cells among gated CD8 + T cells. (B) Frequency of CD8 + T cells (CD45 + B220 − TCRβ + CD8α + ) producing IFN-γ. (C) Total numbers of OVA 257 - or E1 331 -specific IFN-γ + CD8 + T cells (CD45 + B220 − TCRβ + CD8α + IFN-γ + ). (D) Representative dot plots from 10 dpi gated on CD45 + B220 − TCRβ + CD8α + T cells. The numbers embedded in the plots represent the frequency among all gated cells. Polyfunctional cytokine-producing responder CD8 + T cells are classified as IFN-γ single positive (IFN-γ + ), TNF-α single positive (TNF-α + ), or IFN-γ and TNF-α double positive (IFN-γ + TNF-α + ). The pie charts indicate the percentage of each responding population among all responders. (E) Polyfunctional analysis showing the cytokine production and the cytolytic potential of responding CD8 + T cells. Dot plots from 10 dpi are gated on CD45 + B220 − TCRβ + CD8α + T cells, and the numbers embedded in the plots represent the frequency among all gated cells. Responders were classified as IFN-γ + single positive (IFN-γ + ), CD107a single positive (CD107a + ), or IFN-γ + cells that were degranulated (IFN-γ + CD107a + ) in response to OVA 257 or E1 331 peptide stimulation. The pie charts indicate the percentage of each responding population among all responders. (F to H) At 10 dpi, leukocytes were isolated from ipsilateral and contralateral ankle tissue and stimulated with OVA 257 or E1 331 peptide in the presence of brefeldin A for 5 h at 37°C. Mock samples represent cells combined from ipsilateral and contralateral tissues. (F) Representative flow plots from ipsilateral ankle tissue showing IFN-γ production by antigen-specific CD8 + T cells in response to OVA 257 or E1 331 peptide stimulation. The numbers embedded in the flow plots represent the frequency of CD8 + T cells (CD45 + B220 − TCRβ + CD8α + ) producing IFN-γ. (G) Frequency of cells producing IFN-γ among all CD8 + T cells. (H) Total OVA 257 - and E1 331 -specific IFN-γ + CD8 + T cells. The data in the graphs represent the mean ± SEM. Data were combined from 2 to 3 independent experiments. Statistically significant differences between mock- and CHIKV(OVA)-AF-infected mice were determined by one-way analysis of variance with Tukey’s posttest (B and C) or by the Kruskal-Wallis test with Dunn’s posttest (G and H). *, P

Techniques Used: Infection, Mouse Assay, Flow Cytometry, Isolation

Antigen-specific CD8 + T cells accumulate in joint-associated tissue during CHIKV(OVA)-AF infection. WT C57BL/6J mice ( n = 3 to 8 mice/group) were inoculated in the left foot with PBS (mock) or 10 3 PFU of CHIKV(OVA)-AF. At the indicated day postinoculation (dpi), leukocytes were isolated from the ipsilateral (I.) ankle, contralateral (C.) ankle, or spleen and CD8 + T cells were quantified by flow cytometry. (A) Total CD8 + T cells (CD45 + B220 − TCRβ + CD8α + ). (B and C) Frequency (B) and total numbers (C) of CD8 + T cells showing an activated phenotype (CD45 + B220 − TCRβ + CD8α + CD44 hi ). (D and E) Representative flow cytometry plots showing antigen-specific CD8 + T cells in the ipsilateral ankle and contralateral ankle (D) and in spleen (E) of mock-infected or CHIKV(OVA)-AF-infected mice at 14 dpi. The plots are gated on CD45 + B220 − TCRβ + CD8α + cells. The numbers embedded in the plots represent the frequency of K b OVA 257 -positive (K b OVA 257 + ) cells within gated CD8 + T cells. (F) Frequency of K b OVA 257 + cells among activated CD8 + T cells (CD45 + B220 − TCRβ + CD8α + CD44 hi ). (G) Total K b OVA 257 + CD8 + T cells (CD45 + B220 − TCRβ + CD8α + CD44 hi K b OVA 257 + ). The data in the graphs represent the mean ± SEM. Data were combined from 2 to 3 independent experiments. Statistically significant differences from the results for mock-infected mice were determined by one-way analysis of variance with Tukey’s posttest (for panels A, B, and C and for spleen in panels F and G) or by the Kruskal-Wallis test with Dunn’s posttest (for the ipsilateral and contralateral ankles in panels F and G). *, P
Figure Legend Snippet: Antigen-specific CD8 + T cells accumulate in joint-associated tissue during CHIKV(OVA)-AF infection. WT C57BL/6J mice ( n = 3 to 8 mice/group) were inoculated in the left foot with PBS (mock) or 10 3 PFU of CHIKV(OVA)-AF. At the indicated day postinoculation (dpi), leukocytes were isolated from the ipsilateral (I.) ankle, contralateral (C.) ankle, or spleen and CD8 + T cells were quantified by flow cytometry. (A) Total CD8 + T cells (CD45 + B220 − TCRβ + CD8α + ). (B and C) Frequency (B) and total numbers (C) of CD8 + T cells showing an activated phenotype (CD45 + B220 − TCRβ + CD8α + CD44 hi ). (D and E) Representative flow cytometry plots showing antigen-specific CD8 + T cells in the ipsilateral ankle and contralateral ankle (D) and in spleen (E) of mock-infected or CHIKV(OVA)-AF-infected mice at 14 dpi. The plots are gated on CD45 + B220 − TCRβ + CD8α + cells. The numbers embedded in the plots represent the frequency of K b OVA 257 -positive (K b OVA 257 + ) cells within gated CD8 + T cells. (F) Frequency of K b OVA 257 + cells among activated CD8 + T cells (CD45 + B220 − TCRβ + CD8α + CD44 hi ). (G) Total K b OVA 257 + CD8 + T cells (CD45 + B220 − TCRβ + CD8α + CD44 hi K b OVA 257 + ). The data in the graphs represent the mean ± SEM. Data were combined from 2 to 3 independent experiments. Statistically significant differences from the results for mock-infected mice were determined by one-way analysis of variance with Tukey’s posttest (for panels A, B, and C and for spleen in panels F and G) or by the Kruskal-Wallis test with Dunn’s posttest (for the ipsilateral and contralateral ankles in panels F and G). *, P

Techniques Used: Infection, Mouse Assay, Isolation, Flow Cytometry

The viral burdens in joint-associated and lymphoid tissue are similar in CHIKV-infected WT and CD8α −/− mice. WT C57BL/6J mice ( n = 5 to 10 mice/time point) or congenic C57BL/6J CD8α −/− mice ( n = 5 to 9 mice per time point) were inoculated in the left foot with 10 3 PFU of CHIKV-AF. (A to C) On the indicated days postinfection, CHIKV RNA in the ipsilateral (I.) ankle (A), contralateral (C.) ankle (B), and spleen (C) was quantified by RT-qPCR. The data in the graphs represent the mean ± SEM. Data were combined from 4 independent experiments. No statistically significance differences between WT and CD8α −/− mice were detected by two-way analysis of variance.
Figure Legend Snippet: The viral burdens in joint-associated and lymphoid tissue are similar in CHIKV-infected WT and CD8α −/− mice. WT C57BL/6J mice ( n = 5 to 10 mice/time point) or congenic C57BL/6J CD8α −/− mice ( n = 5 to 9 mice per time point) were inoculated in the left foot with 10 3 PFU of CHIKV-AF. (A to C) On the indicated days postinfection, CHIKV RNA in the ipsilateral (I.) ankle (A), contralateral (C.) ankle (B), and spleen (C) was quantified by RT-qPCR. The data in the graphs represent the mean ± SEM. Data were combined from 4 independent experiments. No statistically significance differences between WT and CD8α −/− mice were detected by two-way analysis of variance.

Techniques Used: Infection, Mouse Assay, Quantitative RT-PCR

21) Product Images from "Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2 3"

Article Title: Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2 3

Journal: eNeuro

doi: 10.1523/ENEURO.0020-15.2016

ProBDNF protein levels are elevated acutely after pilocarpine-induced SE in WT C57BL/6J mice. A , Bottom, Representative Western blot of whole hippocampal protein homogenates from WT mice killed 3 h after the induction of SE or time-matched saline controls probed with proBDNF (1:1000) and anti-actin antibodies. Top, Densitometry analysis of proBDNF protein abundance. Ratio of proBDNF/actin at 3 h after SE ( N = 5), expressed as the percentage change relative to mean values (±SEM) of the control group ( N = 5; **** p
Figure Legend Snippet: ProBDNF protein levels are elevated acutely after pilocarpine-induced SE in WT C57BL/6J mice. A , Bottom, Representative Western blot of whole hippocampal protein homogenates from WT mice killed 3 h after the induction of SE or time-matched saline controls probed with proBDNF (1:1000) and anti-actin antibodies. Top, Densitometry analysis of proBDNF protein abundance. Ratio of proBDNF/actin at 3 h after SE ( N = 5), expressed as the percentage change relative to mean values (±SEM) of the control group ( N = 5; **** p

Techniques Used: Mouse Assay, Western Blot

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Generated:

Article Title: Consumptive coagulopathy of severe yellow fever occurs independently of hepatocellular tropism and massive hepatic injury
Article Snippet: .. WT C57BL/6J mice (male and female) were purchased from The Jackson Laboratory. hFRG mice were generated by Yecuris Corporation and maintained according to their specific care and use guidelines. .. In brief, FRG triple knockout mice (Fah −/− , Rag2 −/− , and Il2rɣ −/− on a C57BL/6J background) were fed a diet supplemented orally with nistinone (CurX Nistinone, Yecuris) in the drinking water.

Mouse Assay:

Article Title: Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavag
Article Snippet: .. In WT C57BL/6J mice as well as in HA epitope-tagged BDNF knock-in C57BL/6J mice, there is an increase in proBDNF as early as 3 h after SE onset, with levels remaining elevated at 24 h and peaking at 3 d post-SE. .. The epitope-tagged knock-in C57BL/6J mice were further used to localize early increases in BDNF after SE and HA immunoreactivity was detected primarily in principal cells but also some astrocytes throughout all hippocampal subfields.

Article Title: A High Serum Iron Level Causes Mouse Retinal Iron Accumulation Despite an Intact Blood-Retinal Barrier
Article Snippet: .. WT C57BL/6J mice at 2.5 months were obtained from The Jackson Laboratory (Bar Harbor, ME). .. C57BL/6J mice were treated with or without 1.2 mg of iron sucrose (Venofer; American Regent, Shirley, NY) in 200 mL of 0.9% saline solution via tail vein injection three times (once per week) until sacrifice.

Article Title: Optimal therapeutic activity of monoclonal antibodies against chikungunya virus requires Fc-FcγR interaction on monocytes
Article Snippet: .. Four week-old WT C57BL/6J mice were purchased from Jackson Laboratories. .. Four week-old congenic C1q −/− or FcR common gamma chain deficient ( FcRγ −/− ) mice were bred at the Washington University Animal Facility.

Article Title: The obesity-induced adipokine sST2 exacerbates adipose Treg and ILC2 depletion and promotes insulin resistance
Article Snippet: .. WT C57BL/6J mice (JAX #000664) were purchased from the Jackson Laboratory. .. The Zbtb7b floxed mice were obtained from R. Bosselut from the National Cancer Institute.

Article Title: Consumptive coagulopathy of severe yellow fever occurs independently of hepatocellular tropism and massive hepatic injury
Article Snippet: .. WT C57BL/6J mice (male and female) were purchased from The Jackson Laboratory. hFRG mice were generated by Yecuris Corporation and maintained according to their specific care and use guidelines. .. In brief, FRG triple knockout mice (Fah −/− , Rag2 −/− , and Il2rɣ −/− on a C57BL/6J background) were fed a diet supplemented orally with nistinone (CurX Nistinone, Yecuris) in the drinking water.

Article Title: Generation of Functional Blood Vessels from a Single c-kit+ Adult Vascular Endothelial Stem CellOn the Hunt for Vascular Endothelial Stem Cells
Article Snippet: .. Isolation of Mouse EC Populations Mouse lung ECs were isolated from lungs and other indicated tissues dissected from adult wt C57BL/6J mice (Scanbur AB), GFP-tagged transgenic C57BL/6-Tg(ACTB-EGFP)1Osb/J mice, C57BL/6J-Kdrtm1Jrt; lacZ mice , FVB/N-Tg(TIE2-lacZ)182Sato/J mice , or C57BL/6J-KitW-sh mice (all from The Jackson Laboratory). .. Mouse lung ECs were isolated from lungs dissected from adult mice.

Article Title: Fever Promotes T Lymphocyte Trafficking via a Thermal Sensory Pathway Involving Heat Shock Protein 90 and α4 Integrins
Article Snippet: .. WT C57BL/6J mice were obtained from Jackson Laboratory. .. Itga4 R985A/R985A C57BL/6J mice were generated by Shanghai Biomodel Organism Science & Technology Development Co., Ltd.

Article Title: Rickettsia parkeri Sca2 promotes dissemination in an intradermal infection mouse model
Article Snippet: .. Mouse genotyping Tlr4 −/− , Ifnar −/− , Ifngr −/− , Ifnar −/− Ifngr −/− and WT C57BL/6J mice were previously described and originally obtained from Jackson Laboratories. .. CD-1 mice were obtained from Charles River.

Transgenic Assay:

Article Title: Generation of Functional Blood Vessels from a Single c-kit+ Adult Vascular Endothelial Stem CellOn the Hunt for Vascular Endothelial Stem Cells
Article Snippet: .. Isolation of Mouse EC Populations Mouse lung ECs were isolated from lungs and other indicated tissues dissected from adult wt C57BL/6J mice (Scanbur AB), GFP-tagged transgenic C57BL/6-Tg(ACTB-EGFP)1Osb/J mice, C57BL/6J-Kdrtm1Jrt; lacZ mice , FVB/N-Tg(TIE2-lacZ)182Sato/J mice , or C57BL/6J-KitW-sh mice (all from The Jackson Laboratory). .. Mouse lung ECs were isolated from lungs dissected from adult mice.

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  • 91
    The Jackson Laboratory wt c57bl 6j mice
    ProBDNF protein levels are elevated acutely after pilocarpine-induced SE in WT <t>C57BL/6J</t> mice. A , Bottom, Representative Western blot of whole hippocampal protein homogenates from WT mice killed 3 h after the induction of SE or time-matched saline controls probed with proBDNF (1:1000) and anti-actin antibodies. Top, Densitometry analysis of proBDNF protein abundance. Ratio of proBDNF/actin at 3 h after SE ( N = 5), expressed as the percentage change relative to mean values (±SEM) of the control group ( N = 5; **** p
    Wt C57bl 6j Mice, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 91/100, based on 159 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 91 stars, based on 159 article reviews
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    91
    The Jackson Laboratory wild type c57bl 6j wt
    Effect of Sirt1 overexpression on retinal capillary cell damage in diabetes. Trypsin-digested retinal microvessels from <t>C57BL/6J</t> WT and Sirt1 -overexpressing mice, which had diabetes for ∼8 months, were stained with TUNEL. TUNEL-positive cells were counted throughout the entire retinal vasculature. The microvasculature was then stained with periodic acid Schiff–hematoxylin, providing representative microvasculature ( A ). The arrowhead indicates an acellular capillary, and the arrow points to a pericyte ghost. B : Sirt1 expression was quantified in the retinal cryosections by immunofluorescence (fluoresc) staining using DyLight 488–labeled (green) secondary antibodies. Retinal microvessels prepared with the hypotonic shock method were analyzed for Sirt1 gene expression by SYBR green–based quantitative PCR ( C ) and protein expression by Western blotting ( D ). Values are presented as the mean ± SD ( n = 5–7 mice/group). * P
    Wild Type C57bl 6j Wt, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 91/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    The Jackson Laboratory female wildtype wt c57bl 6j
    Functional validation of Dapp1 as a DEP-responsive gene. A and D) Lung resistance after inhalation exposure to DEP and HDM is not increased in Dapp1 -/- mice compared to WT animals. DEP/HDM exposure induced eosinophilia (B and E) and neutrophilia (C and F) to the same extent in BAL fluid from Dapp1 -/- and WT mice. Mice were first sensitized on day 0 through a 100μl intraperitoneal injection containing 200μg DEP and 25μg HDM + 2.25mg Alum, as an adjuvant. On days 7–10, mice were placed in insulated chambers daily for 20mins supplied with ambient air and saturated with either aerosolized PBS or both 200μg DEP and 25μg HDM. On day 11, airway hyperreactivity was measured by invasive plethysmography, followed by collection of BAL. Cell counts in BAL fluid were determined by flow cytometry. WT control animals were either female <t>C57BL/6J</t> mice purchased from the Jackson Laboratories ( A-C ) or WT littermates of both sexes generated through an intercross between Dapp1 + /- heterozygote mice ( D-F ). For both experiments, n = 4–8 mice in each group. Data are shown as mean ± SE. **** p
    Female Wildtype Wt C57bl 6j, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    The Jackson Laboratory male wt c57bl 6j
    Colitis induction following treatment with anti-CTLA4, but not with anti-CTLA4 DVD. ( A ) Foxp3 IHC staining was performed on GI tract tissue sections from mice receiving different Abs. Arrows indicate Foxp3 + cells. Experiments were conducted twice with 6 mice per group, and the representative figure from each treatment arm was shown. ( B – D ) Recipient C.B17 SCID mice were i.p. injected with 5 × 10 5 purified CD4 + CD45RB hi cells with or without 2.5 × 10 5 CD4 + CD45RB lo cells, and administered the indicated treatments at 200 μg per dose twice per week for 3 weeks. At day 35, all mice were evaluated by endoscopy and scored for colitis. Data were collected from 1 experiment with 10–15 mice per treatment arm. ( B ) Representative endoscopy images of a single mouse from each group. ( C ) Graphic representation of endoscopy scores at day 35. Red dots indicate animals from which colons were further analyzed for percentage of Foxp3 + cells among CD4 + cells in D . ( E – G ) Healthy <t>C57BL/6j</t> mice were injected with either anti-CTLA4 or anti-CTLA4 DVD. ( E ) Flow-gating strategy of spleen samples. ( F ) Percentage of CD44 + ICOS + in CD4 + T cell subsets. ( G ) Percentage of CD44 + ICOS + in CD8 + T cell subsets. For parts E – G , experiments were conducted twice, and the representative data from 1 experiment was from 4 mice per group. Bars represent mean or mean ± SEM. * P
    Male Wt C57bl 6j, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ProBDNF protein levels are elevated acutely after pilocarpine-induced SE in WT C57BL/6J mice. A , Bottom, Representative Western blot of whole hippocampal protein homogenates from WT mice killed 3 h after the induction of SE or time-matched saline controls probed with proBDNF (1:1000) and anti-actin antibodies. Top, Densitometry analysis of proBDNF protein abundance. Ratio of proBDNF/actin at 3 h after SE ( N = 5), expressed as the percentage change relative to mean values (±SEM) of the control group ( N = 5; **** p

    Journal: eNeuro

    Article Title: Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2Rapid Increases in proBDNF after Pilocarpine-Induced Status Epilepticus in Mice Are Associated with Reduced proBDNF Cleavage Machinery 1 2 3

    doi: 10.1523/ENEURO.0020-15.2016

    Figure Lengend Snippet: ProBDNF protein levels are elevated acutely after pilocarpine-induced SE in WT C57BL/6J mice. A , Bottom, Representative Western blot of whole hippocampal protein homogenates from WT mice killed 3 h after the induction of SE or time-matched saline controls probed with proBDNF (1:1000) and anti-actin antibodies. Top, Densitometry analysis of proBDNF protein abundance. Ratio of proBDNF/actin at 3 h after SE ( N = 5), expressed as the percentage change relative to mean values (±SEM) of the control group ( N = 5; **** p

    Article Snippet: In WT C57BL/6J mice as well as in HA epitope-tagged BDNF knock-in C57BL/6J mice, there is an increase in proBDNF as early as 3 h after SE onset, with levels remaining elevated at 24 h and peaking at 3 d post-SE.

    Techniques: Mouse Assay, Western Blot

    Effect of Sirt1 overexpression on retinal capillary cell damage in diabetes. Trypsin-digested retinal microvessels from C57BL/6J WT and Sirt1 -overexpressing mice, which had diabetes for ∼8 months, were stained with TUNEL. TUNEL-positive cells were counted throughout the entire retinal vasculature. The microvasculature was then stained with periodic acid Schiff–hematoxylin, providing representative microvasculature ( A ). The arrowhead indicates an acellular capillary, and the arrow points to a pericyte ghost. B : Sirt1 expression was quantified in the retinal cryosections by immunofluorescence (fluoresc) staining using DyLight 488–labeled (green) secondary antibodies. Retinal microvessels prepared with the hypotonic shock method were analyzed for Sirt1 gene expression by SYBR green–based quantitative PCR ( C ) and protein expression by Western blotting ( D ). Values are presented as the mean ± SD ( n = 5–7 mice/group). * P

    Journal: Diabetes

    Article Title: Sirt1: A Guardian of the Development of Diabetic Retinopathy

    doi: 10.2337/db17-0996

    Figure Lengend Snippet: Effect of Sirt1 overexpression on retinal capillary cell damage in diabetes. Trypsin-digested retinal microvessels from C57BL/6J WT and Sirt1 -overexpressing mice, which had diabetes for ∼8 months, were stained with TUNEL. TUNEL-positive cells were counted throughout the entire retinal vasculature. The microvasculature was then stained with periodic acid Schiff–hematoxylin, providing representative microvasculature ( A ). The arrowhead indicates an acellular capillary, and the arrow points to a pericyte ghost. B : Sirt1 expression was quantified in the retinal cryosections by immunofluorescence (fluoresc) staining using DyLight 488–labeled (green) secondary antibodies. Retinal microvessels prepared with the hypotonic shock method were analyzed for Sirt1 gene expression by SYBR green–based quantitative PCR ( C ) and protein expression by Western blotting ( D ). Values are presented as the mean ± SD ( n = 5–7 mice/group). * P

    Article Snippet: Mice Diabetes was induced in wild-type C57BL/6J (WT) and Sirt1 -overexpressing (St , C57BL/6-Actbtm3.1 [Sirt1 ] Npa/J ; Sirt1 ) mice (The Jackson Laboratory, Bar Harbor, ME), with a body weight (BW) of ∼20 g (either sex), by streptozotocin injection (55 mg/kg BW for four consecutive days).

    Techniques: Over Expression, Mouse Assay, Staining, TUNEL Assay, Expressing, Immunofluorescence, Labeling, SYBR Green Assay, Real-time Polymerase Chain Reaction, Western Blot

    Functional validation of Dapp1 as a DEP-responsive gene. A and D) Lung resistance after inhalation exposure to DEP and HDM is not increased in Dapp1 -/- mice compared to WT animals. DEP/HDM exposure induced eosinophilia (B and E) and neutrophilia (C and F) to the same extent in BAL fluid from Dapp1 -/- and WT mice. Mice were first sensitized on day 0 through a 100μl intraperitoneal injection containing 200μg DEP and 25μg HDM + 2.25mg Alum, as an adjuvant. On days 7–10, mice were placed in insulated chambers daily for 20mins supplied with ambient air and saturated with either aerosolized PBS or both 200μg DEP and 25μg HDM. On day 11, airway hyperreactivity was measured by invasive plethysmography, followed by collection of BAL. Cell counts in BAL fluid were determined by flow cytometry. WT control animals were either female C57BL/6J mice purchased from the Jackson Laboratories ( A-C ) or WT littermates of both sexes generated through an intercross between Dapp1 + /- heterozygote mice ( D-F ). For both experiments, n = 4–8 mice in each group. Data are shown as mean ± SE. **** p

    Journal: PLoS Genetics

    Article Title: A GWAS approach identifies Dapp1 as a determinant of air pollution-induced airway hyperreactivity

    doi: 10.1371/journal.pgen.1008528

    Figure Lengend Snippet: Functional validation of Dapp1 as a DEP-responsive gene. A and D) Lung resistance after inhalation exposure to DEP and HDM is not increased in Dapp1 -/- mice compared to WT animals. DEP/HDM exposure induced eosinophilia (B and E) and neutrophilia (C and F) to the same extent in BAL fluid from Dapp1 -/- and WT mice. Mice were first sensitized on day 0 through a 100μl intraperitoneal injection containing 200μg DEP and 25μg HDM + 2.25mg Alum, as an adjuvant. On days 7–10, mice were placed in insulated chambers daily for 20mins supplied with ambient air and saturated with either aerosolized PBS or both 200μg DEP and 25μg HDM. On day 11, airway hyperreactivity was measured by invasive plethysmography, followed by collection of BAL. Cell counts in BAL fluid were determined by flow cytometry. WT control animals were either female C57BL/6J mice purchased from the Jackson Laboratories ( A-C ) or WT littermates of both sexes generated through an intercross between Dapp1 + /- heterozygote mice ( D-F ). For both experiments, n = 4–8 mice in each group. Data are shown as mean ± SE. **** p

    Article Snippet: We first used the same exposure protocol as for the HMDP strain survey ( ) to characterize female Dapp1 -/- mice and female wildtype (WT) C57BL/6J controls purchased from the Jackson Laboratories.

    Techniques: Functional Assay, Mouse Assay, Injection, Flow Cytometry, Cytometry, Generated

    Colitis induction following treatment with anti-CTLA4, but not with anti-CTLA4 DVD. ( A ) Foxp3 IHC staining was performed on GI tract tissue sections from mice receiving different Abs. Arrows indicate Foxp3 + cells. Experiments were conducted twice with 6 mice per group, and the representative figure from each treatment arm was shown. ( B – D ) Recipient C.B17 SCID mice were i.p. injected with 5 × 10 5 purified CD4 + CD45RB hi cells with or without 2.5 × 10 5 CD4 + CD45RB lo cells, and administered the indicated treatments at 200 μg per dose twice per week for 3 weeks. At day 35, all mice were evaluated by endoscopy and scored for colitis. Data were collected from 1 experiment with 10–15 mice per treatment arm. ( B ) Representative endoscopy images of a single mouse from each group. ( C ) Graphic representation of endoscopy scores at day 35. Red dots indicate animals from which colons were further analyzed for percentage of Foxp3 + cells among CD4 + cells in D . ( E – G ) Healthy C57BL/6j mice were injected with either anti-CTLA4 or anti-CTLA4 DVD. ( E ) Flow-gating strategy of spleen samples. ( F ) Percentage of CD44 + ICOS + in CD4 + T cell subsets. ( G ) Percentage of CD44 + ICOS + in CD8 + T cell subsets. For parts E – G , experiments were conducted twice, and the representative data from 1 experiment was from 4 mice per group. Bars represent mean or mean ± SEM. * P

    Journal: The Journal of Clinical Investigation

    Article Title: Tumor-conditional anti-CTLA4 uncouples antitumor efficacy from immunotherapy-related toxicity

    doi: 10.1172/JCI123391

    Figure Lengend Snippet: Colitis induction following treatment with anti-CTLA4, but not with anti-CTLA4 DVD. ( A ) Foxp3 IHC staining was performed on GI tract tissue sections from mice receiving different Abs. Arrows indicate Foxp3 + cells. Experiments were conducted twice with 6 mice per group, and the representative figure from each treatment arm was shown. ( B – D ) Recipient C.B17 SCID mice were i.p. injected with 5 × 10 5 purified CD4 + CD45RB hi cells with or without 2.5 × 10 5 CD4 + CD45RB lo cells, and administered the indicated treatments at 200 μg per dose twice per week for 3 weeks. At day 35, all mice were evaluated by endoscopy and scored for colitis. Data were collected from 1 experiment with 10–15 mice per treatment arm. ( B ) Representative endoscopy images of a single mouse from each group. ( C ) Graphic representation of endoscopy scores at day 35. Red dots indicate animals from which colons were further analyzed for percentage of Foxp3 + cells among CD4 + cells in D . ( E – G ) Healthy C57BL/6j mice were injected with either anti-CTLA4 or anti-CTLA4 DVD. ( E ) Flow-gating strategy of spleen samples. ( F ) Percentage of CD44 + ICOS + in CD4 + T cell subsets. ( G ) Percentage of CD44 + ICOS + in CD8 + T cell subsets. For parts E – G , experiments were conducted twice, and the representative data from 1 experiment was from 4 mice per group. Bars represent mean or mean ± SEM. * P

    Article Snippet: Eight- to ten-week-old male WT C57BL/6j and Rag 1–/– mice (on C57BL/6j background) were purchased from Jackson Laboratory for the following mouse experiments.

    Techniques: Immunohistochemistry, Staining, Mouse Assay, Injection, Purification, Flow Cytometry

    Immune landscape within tumors after different CTLA4 blockades. C57BL/6j male mice were implanted with TRAMP-C2 tumors at day 0, and tumors were allowed to grow for 40 days. Mice were randomized into different treatment groups and received different treatments at days 40, 43, 46, and 49. Tissues were harvested at day 52 and analyzed by flow cytometry. ( A ) Tumors were harvested from each treatment group and analyzed for different immune subsets by flow cytometry. ( B ) Total numbers of CD4 + T cells per tumor weight. CD4 + T cells were pregated on live CD45 + CD3 + CD8 – T cells. ( C ) Percentage of ICOS expression among CD4 + Foxp3 – T cells. ( D ) Percentage of CD4 + Foxp3 + among CD3 + cells. ( E ) Total number of infiltrating CD8 + T cells per tumor weight. ( F ) Total numbers of CD8 + CD25 + T cells per tumor weight. ( G ) Percentage of Ki-67 among CD3 + CD8 + T cells. ( H ) Percentage of Spas-1–reactive CD8 + T cells among CD3 + cells. Two independent experiments were conducted. Data were shown as 5 mice per group from 1 representative experiment. Bars represent mean ± SEM. * P

    Journal: The Journal of Clinical Investigation

    Article Title: Tumor-conditional anti-CTLA4 uncouples antitumor efficacy from immunotherapy-related toxicity

    doi: 10.1172/JCI123391

    Figure Lengend Snippet: Immune landscape within tumors after different CTLA4 blockades. C57BL/6j male mice were implanted with TRAMP-C2 tumors at day 0, and tumors were allowed to grow for 40 days. Mice were randomized into different treatment groups and received different treatments at days 40, 43, 46, and 49. Tissues were harvested at day 52 and analyzed by flow cytometry. ( A ) Tumors were harvested from each treatment group and analyzed for different immune subsets by flow cytometry. ( B ) Total numbers of CD4 + T cells per tumor weight. CD4 + T cells were pregated on live CD45 + CD3 + CD8 – T cells. ( C ) Percentage of ICOS expression among CD4 + Foxp3 – T cells. ( D ) Percentage of CD4 + Foxp3 + among CD3 + cells. ( E ) Total number of infiltrating CD8 + T cells per tumor weight. ( F ) Total numbers of CD8 + CD25 + T cells per tumor weight. ( G ) Percentage of Ki-67 among CD3 + CD8 + T cells. ( H ) Percentage of Spas-1–reactive CD8 + T cells among CD3 + cells. Two independent experiments were conducted. Data were shown as 5 mice per group from 1 representative experiment. Bars represent mean ± SEM. * P

    Article Snippet: Eight- to ten-week-old male WT C57BL/6j and Rag 1–/– mice (on C57BL/6j background) were purchased from Jackson Laboratory for the following mouse experiments.

    Techniques: Mouse Assay, Flow Cytometry, Cytometry, Expressing

    Anti-CTLA4 DVD reduced treatment-associated toxicity in mice. Eight- to ten-week-old male Rag1 –/– mice (on a C57BL/6j background) were adoptively transferred with purified CD4 + CD25 – CD45RB hi cells from WT mice. Recipient mice were then treated with either anti-CTLA4, anti-CTLA4 DVD, or isotype control. ( A ) Percentage of body weight lost among different treatment groups. ( B ) Mice were harvested at day 28, and multiorgan pathology was examined among different treatment groups. Arrows represent lymphocytic infiltration in the tissues. Experiments were conducted twice, and data shown are from 5 mice per group. Bars represent mean ± SEM. ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Tumor-conditional anti-CTLA4 uncouples antitumor efficacy from immunotherapy-related toxicity

    doi: 10.1172/JCI123391

    Figure Lengend Snippet: Anti-CTLA4 DVD reduced treatment-associated toxicity in mice. Eight- to ten-week-old male Rag1 –/– mice (on a C57BL/6j background) were adoptively transferred with purified CD4 + CD25 – CD45RB hi cells from WT mice. Recipient mice were then treated with either anti-CTLA4, anti-CTLA4 DVD, or isotype control. ( A ) Percentage of body weight lost among different treatment groups. ( B ) Mice were harvested at day 28, and multiorgan pathology was examined among different treatment groups. Arrows represent lymphocytic infiltration in the tissues. Experiments were conducted twice, and data shown are from 5 mice per group. Bars represent mean ± SEM. ** P

    Article Snippet: Eight- to ten-week-old male WT C57BL/6j and Rag 1–/– mice (on C57BL/6j background) were purchased from Jackson Laboratory for the following mouse experiments.

    Techniques: Mouse Assay, Purification

    Anti-CTLA4 DVD preserves antitumor activity. ( A ) C57BL/6j male mice were implanted with TRAMP-C2 tumors at day 0 (D0), and tumors were allowed to grow for 40 days. Mice were randomized into different treatment groups before Ab injection and then received different treatments at days 40, 43, 46, and 49. ( B and C ) Tumor growth from different treatment groups over time. ( D ) Survival curve of different treatment groups. Data were conducted with 2 independent experiments. Each treatment arm was collected from 7 mice per group. Bars represent mean ± SEM. * P

    Journal: The Journal of Clinical Investigation

    Article Title: Tumor-conditional anti-CTLA4 uncouples antitumor efficacy from immunotherapy-related toxicity

    doi: 10.1172/JCI123391

    Figure Lengend Snippet: Anti-CTLA4 DVD preserves antitumor activity. ( A ) C57BL/6j male mice were implanted with TRAMP-C2 tumors at day 0 (D0), and tumors were allowed to grow for 40 days. Mice were randomized into different treatment groups before Ab injection and then received different treatments at days 40, 43, 46, and 49. ( B and C ) Tumor growth from different treatment groups over time. ( D ) Survival curve of different treatment groups. Data were conducted with 2 independent experiments. Each treatment arm was collected from 7 mice per group. Bars represent mean ± SEM. * P

    Article Snippet: Eight- to ten-week-old male WT C57BL/6j and Rag 1–/– mice (on C57BL/6j background) were purchased from Jackson Laboratory for the following mouse experiments.

    Techniques: Activity Assay, Mouse Assay, Injection

    Anti-CTLA4–mediated immune-related toxicities in a murine model. Eight- to ten-week-old male Rag 1 –/– mice (on C57BL/6j background) were adoptively transferred with purified CD4 + CD25 – CD45RB hi cells from WT mice. ( A ) Sorting strategy. ( B ) Body weight loss over time after treatment. ( C ) Different organs were harvested at day 45, and pathological changes were examined using H E staining under a microscope. Arrows indicate lymphocytic infiltration and pathological changes. ( D – F ) Target organ pathological scores were evaluated by a board-certified pathologist in a single-blind fashion. ( G ) Splenocytes were harvested at day 45 from different treatment groups, and CD4 + T cells were examined for TNF-α secretion by flow cytometry. ( H ) Percentage of TNF-α + CD4 + T cells among CD45 + cells. Experiments were conducted twice, and data were shown with 5 mice per group. Three mice from the Rag1 –/– only group were used as negative control. For the TNF-α experiment, data were collected from 3 mice per group. Bars represent mean ± SEM. * P

    Journal: The Journal of Clinical Investigation

    Article Title: Tumor-conditional anti-CTLA4 uncouples antitumor efficacy from immunotherapy-related toxicity

    doi: 10.1172/JCI123391

    Figure Lengend Snippet: Anti-CTLA4–mediated immune-related toxicities in a murine model. Eight- to ten-week-old male Rag 1 –/– mice (on C57BL/6j background) were adoptively transferred with purified CD4 + CD25 – CD45RB hi cells from WT mice. ( A ) Sorting strategy. ( B ) Body weight loss over time after treatment. ( C ) Different organs were harvested at day 45, and pathological changes were examined using H E staining under a microscope. Arrows indicate lymphocytic infiltration and pathological changes. ( D – F ) Target organ pathological scores were evaluated by a board-certified pathologist in a single-blind fashion. ( G ) Splenocytes were harvested at day 45 from different treatment groups, and CD4 + T cells were examined for TNF-α secretion by flow cytometry. ( H ) Percentage of TNF-α + CD4 + T cells among CD45 + cells. Experiments were conducted twice, and data were shown with 5 mice per group. Three mice from the Rag1 –/– only group were used as negative control. For the TNF-α experiment, data were collected from 3 mice per group. Bars represent mean ± SEM. * P

    Article Snippet: Eight- to ten-week-old male WT C57BL/6j and Rag 1–/– mice (on C57BL/6j background) were purchased from Jackson Laboratory for the following mouse experiments.

    Techniques: Mouse Assay, Purification, Staining, Microscopy, Flow Cytometry, Cytometry, Negative Control