c57bl 6 wt mice (Charles River Laboratories)

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Charles River Laboratories c57bl 6 wt mice
Herpes simplex virus 1 replication in the brains of TRIF −/− , IPS-1 −/− , and <t>C57BL/6</t> WT mice. (A) Viral DNA loads were measured in brain homogenates by qPCR. Results are expressed as copies/ng of extracted DNA. (B) Viral

Herpes simplex virus 1 replication in the brains of TRIF −/− , IPS-1 −/− , and <t>C57BL/6</t> WT mice. (A) Viral DNA loads were measured in brain homogenates by qPCR. Results are expressed as copies/ng of extracted DNA. (B) Viral

C57bl 6 Wt Mice, supplied by Charles River Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 104 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Both TRIF and IPS-1 Adaptor Proteins Contribute to the Cerebral Innate Immune Response against Herpes Simplex Virus 1 Infection"

Article Title: Both TRIF and IPS-1 Adaptor Proteins Contribute to the Cerebral Innate Immune Response against Herpes Simplex Virus 1 Infection

Journal: Journal of Virology

doi: 10.1128/JVI.00591-13

Herpes simplex virus 1 replication in the brains of TRIF −/− , IPS-1 −/− , and C57BL/6 WT mice. (A) Viral DNA loads were measured in brain homogenates by qPCR. Results are expressed as copies/ng of extracted DNA. (B) Viral

Figure Legend Snippet: Herpes simplex virus 1 replication in the brains of TRIF −/− , IPS-1 −/− , and C57BL/6 WT mice. (A) Viral DNA loads were measured in brain homogenates by qPCR. Results are expressed as copies/ng of extracted DNA. (B) Viral

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

Levels of phosphorylated IRF-3 (A, B, and C) or IRF-7 (D, E, and F) in C57BL/6 WT (A and D), TRIF −/− (B and E), or IPS-1 −/− (C and F) mice. Animals were infected intranasally with 7.5 × 10 5 PFU of HSV-1. Proteins

Figure Legend Snippet: Levels of phosphorylated IRF-3 (A, B, and C) or IRF-7 (D, E, and F) in C57BL/6 WT (A and D), TRIF −/− (B and E), or IPS-1 −/− (C and F) mice. Animals were infected intranasally with 7.5 × 10 5 PFU of HSV-1. Proteins

Techniques Used: Mouse Assay, Infection

Swollen forehead of HSV-1-infected mice on day 7 postinfection (A) and survival rates of TRIF −/− (B) or IPS-1 −/− (C) mice compared to those of C57BL/6 WT mice. Ten to fourteen animals per group were infected intranasally

Figure Legend Snippet: Swollen forehead of HSV-1-infected mice on day 7 postinfection (A) and survival rates of TRIF −/− (B) or IPS-1 −/− (C) mice compared to those of C57BL/6 WT mice. Ten to fourteen animals per group were infected intranasally

Techniques Used: Infection, Mouse Assay

Levels of IFN-β (A) or IFN-α (B) in brain homogenates from TRIF −/− , IPS-1 −/− , and C57BL/6 WT mice. IFN-α/β levels (expressed in pg/ml) were measured in brain homogenate supernatants by ELISA

Figure Legend Snippet: Levels of IFN-β (A) or IFN-α (B) in brain homogenates from TRIF −/− , IPS-1 −/− , and C57BL/6 WT mice. IFN-α/β levels (expressed in pg/ml) were measured in brain homogenate supernatants by ELISA

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

2) Product Images from "Conditional Expression of Human 15-Lipoxygenase-1 in Mouse Prostate Induces Prostatic Intraepithelial Neoplasia: The FLiMP Mouse Model 1"

Article Title: Conditional Expression of Human 15-Lipoxygenase-1 in Mouse Prostate Induces Prostatic Intraepithelial Neoplasia: The FLiMP Mouse Model 1

Journal: Neoplasia (New York, N.Y.)

doi:

Real-time qRT-PCR screening. Extracted RNA were quantitated for mouse β-actin and h15-LO-1 from dissected prostate lobes of wt age-matched C57BL/6 littermate controls (A) compared to hemizygous (B) and homozygous (C) FLiMP mice. RNA from dissected dorsolateral, anterior, and ventral prostate lobes, each from three transgenic C57BL/6 FLiMP +/- (hemizygous) and FLiMP +/+ (homozygous) mice at 10 weeks of age, were compared to control RNA similarly extracted from wt mice. Real-time quantitations were performed using the iQ5 Real-Time PCR Detection System, as described in the Materials and Methods section. A comparative relative expression level of h15-LO-1 mRNA in different prostate lobes of homozygous (□) and hemizygous ( ) mice is represented (D). Fluorescence threshold values were calculated using the system software. (◆) 15-LO-1 anterior; (■) 15-LO-1 lateral; (▴) 15-LO-1 ventral; (x) 15-LO-1 dorsal; (*) β-actin anterior; (●) β-actin lateral; (+) β-actin ventral; (-) β-actin dorsal.

Figure Legend Snippet: Real-time qRT-PCR screening. Extracted RNA were quantitated for mouse β-actin and h15-LO-1 from dissected prostate lobes of wt age-matched C57BL/6 littermate controls (A) compared to hemizygous (B) and homozygous (C) FLiMP mice. RNA from dissected dorsolateral, anterior, and ventral prostate lobes, each from three transgenic C57BL/6 FLiMP +/- (hemizygous) and FLiMP +/+ (homozygous) mice at 10 weeks of age, were compared to control RNA similarly extracted from wt mice. Real-time quantitations were performed using the iQ5 Real-Time PCR Detection System, as described in the Materials and Methods section. A comparative relative expression level of h15-LO-1 mRNA in different prostate lobes of homozygous (□) and hemizygous ( ) mice is represented (D). Fluorescence threshold values were calculated using the system software. (◆) 15-LO-1 anterior; (■) 15-LO-1 lateral; (▴) 15-LO-1 ventral; (x) 15-LO-1 dorsal; (*) β-actin anterior; (●) β-actin lateral; (+) β-actin ventral; (-) β-actin dorsal.

Techniques Used: Quantitative RT-PCR, Mouse Assay, Transgenic Assay, Real-time Polymerase Chain Reaction, Expressing, Fluorescence, Software

13-HODE levels in tissues, as a measure of 15-LO-1 activity, were examined using commercially available ELISA plates, as described in Materials and Methods section. Concentrations of 13-(S)-HODE were represented as micrograms per milliliter per gram of tissue after normalizing for the endogenous murine 12/15-LO protein similarly extracted from transgenic wt littermate control samples. Proteins extracted from dissected dorsolateral ( ), ventral (□), and anterior (■) prostate lobes, each from three transgenic C57BL/6, FLiMP +/- (A), and FLiMP +/+ (B) mice at 8, 14, 21, and 32 weeks of age, were compared to age-matched nontransgenic C57BL/6 littermate wt control proteins that were similarly extracted and pooled.

Figure Legend Snippet: 13-HODE levels in tissues, as a measure of 15-LO-1 activity, were examined using commercially available ELISA plates, as described in Materials and Methods section. Concentrations of 13-(S)-HODE were represented as micrograms per milliliter per gram of tissue after normalizing for the endogenous murine 12/15-LO protein similarly extracted from transgenic wt littermate control samples. Proteins extracted from dissected dorsolateral ( ), ventral (□), and anterior (■) prostate lobes, each from three transgenic C57BL/6, FLiMP +/- (A), and FLiMP +/+ (B) mice at 8, 14, 21, and 32 weeks of age, were compared to age-matched nontransgenic C57BL/6 littermate wt control proteins that were similarly extracted and pooled.

Techniques Used: Activity Assay, Enzyme-linked Immunosorbent Assay, Transgenic Assay, Mouse Assay

3) Product Images from "Dendritic cell functional improvement in a preclinical model of lentiviral-mediated gene therapy for Wiskott-Aldrich syndrome"

Article Title: Dendritic cell functional improvement in a preclinical model of lentiviral-mediated gene therapy for Wiskott-Aldrich syndrome

Journal: Gene therapy

doi: 10.1038/gt.2011.202

Figure Legend Snippet: BMDCs of GT mice rescue T cell priming capacity. ( a-b ) Five ×10 5 BMDCs of BMT wt, BMT was −/− or GT mice were loaded with 0.05 nM of SIINFEKL OVA peptide and injected subcutaneously in C57BL/6 (CD45.2) wt recipient mice that had been transferred with 1.5×10 6 CFSE-labeled OT-I/CD45.1 cells 24 hours before. Draining LNs were collected 3 days after and T cell proliferation was analyzed by flow cytometry. ( a ) Representative histograms showing CFSE dilution profile of transferred OT-I cells, gated on CD8 + /CD45.1 + live cells. ( b ) Quantification of T cell expansion expressed as percentage of OT-I cells over the total CD8 + T cell population. Each symbol represents an individual mouse; horizontal lines indicate mean values. ( c-e ) Five ×10 5 BMDCs of BMT wt, BMT was −/− or GT mice were loaded with OVA (150 μg/ml) and injected subcutaneously in C57BL/6 (CD45.1) wt recipient mice that had been transferred with 1.5×10 6 CFSE-labeled OT-II/CD45.2 cells 24 hours before. Draining LNs were collected 3 days later and T cell proliferation and intracellular IFN-γ production was analyzed by flow cytometry. ( c ) Representative histograms showing CFSE dilution profile of transferred OT-II cells, gated on CD4 + /CD45.2 + live cells. ( d ) Data obtained from the analysis of CFSE dilution were expressed as the percentage of OT-II cells that remain undivided, that divided one to four times, or that divided five to eight times. ( e ) Quantification of T cell expansion expressed as percentage of OT-II cells over the total CD4 + T cell population. ( f ) Percentage of IFN-γ expressing OT-II cells, evaluated by flow cytometry. Each symbol represents an individual mouse; horizontal lines indicate mean values (n.s., not significant; *p

Techniques Used: Mouse Assay, Injection, Labeling, Flow Cytometry, Cytometry, Expressing

Amelioration of in vivo migration of BMDCs of GT mice. ( a ) 0.5-2×10 6 CFSE-labeled BMDCs from BMT wt, BMT was −/− and GT mice were injected into the footpad of C57BL/6 wt mice. Single cell suspensions of popliteal LNs were stained for CD11c and analyzed by flow cytometry. Numbers represent CFSE + /CD11c + cells, gated on live cells. ( b ) Numbers of BMT wt, BMT was −/− and GT migrated DCs were normalized to BMT wt migrated DCs and expressed as percentage. BMT wt (black); BMT was −/− (white); GT mice (gray). Bars represent the means + SEM of three different experiments with three mice per group (n.s., not significant; *p

Figure Legend Snippet: Amelioration of in vivo migration of BMDCs of GT mice. ( a ) 0.5-2×10 6 CFSE-labeled BMDCs from BMT wt, BMT was −/− and GT mice were injected into the footpad of C57BL/6 wt mice. Single cell suspensions of popliteal LNs were stained for CD11c and analyzed by flow cytometry. Numbers represent CFSE + /CD11c + cells, gated on live cells. ( b ) Numbers of BMT wt, BMT was −/− and GT migrated DCs were normalized to BMT wt migrated DCs and expressed as percentage. BMT wt (black); BMT was −/− (white); GT mice (gray). Bars represent the means + SEM of three different experiments with three mice per group (n.s., not significant; *p

Techniques Used: In Vivo, Migration, Mouse Assay, Labeling, Injection, Staining, Flow Cytometry, Cytometry

4) Product Images from "Essential role for histone deacetylase 11 (HDAC11) in neutrophil biology"

Article Title: Essential role for histone deacetylase 11 (HDAC11) in neutrophil biology

Journal: Journal of Leukocyte Biology

doi: 10.1189/jlb.1A0415-176RRR

Septic shock experiment comparing HDAC11KO with C56BL/6 mice. A cohort ( n = 8) of HDAC11KO and C57BL/6 mice was injected with 15 mg/kg LPS via TV. Survival graph representing both groups in timeline up to 72 h (data presented are representative of 2 individual experiments).

Figure Legend Snippet: Septic shock experiment comparing HDAC11KO with C56BL/6 mice. A cohort ( n = 8) of HDAC11KO and C57BL/6 mice was injected with 15 mg/kg LPS via TV. Survival graph representing both groups in timeline up to 72 h (data presented are representative of 2 individual experiments).

Techniques Used: Mouse Assay, Injection

Observation of splenomegaly as well as hypercellularity with granulocytic expansion in BM of HDAC11KO mice. (A) Representative sections of spleens (H E stain) harvested from aged C57BL/6 WT (upper) and HDAC11 KO (lower) mice showing a marked expansion of the red pulp (left; original magnification, ×40), as a result of replacement of the mostly lymphocytic red pulp cellularity observed on C57BL/6 WT mice with mostly trilineage extramedullary hematopoiesis on HDAC11KO mice (right; original magnification, ×400). (B) Representative sections of femurs (H E stain) harvested from aged C57BL/6 WT (upper) and HDAC11 KO (lower) mice showing a marked increase in the BM cellularity on HDAC11KO mice compared with WT mice (left; original magnification, ×20), mostly as a result of an expansion on maturing neutrophils (right; original magnification, ×200; n = 5/group).

Figure Legend Snippet: Observation of splenomegaly as well as hypercellularity with granulocytic expansion in BM of HDAC11KO mice. (A) Representative sections of spleens (H E stain) harvested from aged C57BL/6 WT (upper) and HDAC11 KO (lower) mice showing a marked expansion of the red pulp (left; original magnification, ×40), as a result of replacement of the mostly lymphocytic red pulp cellularity observed on C57BL/6 WT mice with mostly trilineage extramedullary hematopoiesis on HDAC11KO mice (right; original magnification, ×400). (B) Representative sections of femurs (H E stain) harvested from aged C57BL/6 WT (upper) and HDAC11 KO (lower) mice showing a marked increase in the BM cellularity on HDAC11KO mice compared with WT mice (left; original magnification, ×20), mostly as a result of an expansion on maturing neutrophils (right; original magnification, ×200; n = 5/group).

Techniques Used: Mouse Assay, H&E Stain

5) Product Images from "Cytomegalovirus-Specific IL-10-Producing CD4+ T Cells Are Governed by Type-I IFN-Induced IL-27 and Promote Virus Persistence"

Article Title: Cytomegalovirus-Specific IL-10-Producing CD4+ T Cells Are Governed by Type-I IFN-Induced IL-27 and Promote Virus Persistence

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1006050

Type-I IFN induces IL-27 expression during MCMV infection. (A) Heat map of IL-27 p28 mRNA expression by WT, IFNβ -/- and IFNαR -/- macrophages following stimulation with WT MCMV (C3X), replication-deficient (ΔIE3) MCMV or poly(I:C). (B) Expression of IL-27 p28 by splenic leukocytes was assessed d2 pi in Cre - mice treated/not with 2 mg anti-IFNαR-1. Mean + SEM of 8 mice/group is shown and data is representative of 2 separate experiments. (C) WT (C57BL/6) mice were infected with MCMV treated with anti-IL-27, Isotype or virus alone. At d6 pi mice were treated /not with anti-IL-10R and at d14 pi spleen CD4 + /IFNγ + (D) responses were quantified and expressed as mean ± SEM of 4–15 mice/group. (E) MCMV genomes in saliva were quantified by qPCR. Data shown as mean ± SEM from 4–15 mice/ group.

Figure Legend Snippet: Type-I IFN induces IL-27 expression during MCMV infection. (A) Heat map of IL-27 p28 mRNA expression by WT, IFNβ -/- and IFNαR -/- macrophages following stimulation with WT MCMV (C3X), replication-deficient (ΔIE3) MCMV or poly(I:C). (B) Expression of IL-27 p28 by splenic leukocytes was assessed d2 pi in Cre - mice treated/not with 2 mg anti-IFNαR-1. Mean + SEM of 8 mice/group is shown and data is representative of 2 separate experiments. (C) WT (C57BL/6) mice were infected with MCMV treated with anti-IL-27, Isotype or virus alone. At d6 pi mice were treated /not with anti-IL-10R and at d14 pi spleen CD4 + /IFNγ + (D) responses were quantified and expressed as mean ± SEM of 4–15 mice/group. (E) MCMV genomes in saliva were quantified by qPCR. Data shown as mean ± SEM from 4–15 mice/ group.

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

IL-27 promotes splenic CD4 + IL-10 + T cell development whereas ICOS is required for salivary gland CD4 + IL-10 + T cell accumulation. Il-27r α -/- or WT (C57BL/6) mice were infected with MCMV and spleen and salivary gland CD4 + /IL-10 + (A) responses were quantified and expressed at mean + SEM of 11 mice/group. (B) Representative bivariant FACS plots of IFNγ versus IL-10 expression by splenic CD4 + CD3 + T cells. (C D) gp130 and (E) ICOS expression by IL-10 + (Thy1.1 + ) and IL-10 - (Thy1.1 - ) CD4 + CD3 + T cells was assessed in 10-Bit mice and shown as representative FACS plots (C) and histogram overlays (E) with mean + SEM of 5–6 mice/group (D). Gating was determined using Thy1.1 + CD4 + CD3 + cells derived from fluorescent minus one-stained samples from mice infected for 14 days. (F G) WT (C57BL/6) mice were infected with MCMV and at d6 and d10 pi αICOS or Isotype antibody was added. At d14 pi (F) salivary glands and (G) spleen CD4 + /IL-10 + responses were quantified and expressed as mean + SEM of 6 mice/group.

Figure Legend Snippet: IL-27 promotes splenic CD4 + IL-10 + T cell development whereas ICOS is required for salivary gland CD4 + IL-10 + T cell accumulation. Il-27r α -/- or WT (C57BL/6) mice were infected with MCMV and spleen and salivary gland CD4 + /IL-10 + (A) responses were quantified and expressed at mean + SEM of 11 mice/group. (B) Representative bivariant FACS plots of IFNγ versus IL-10 expression by splenic CD4 + CD3 + T cells. (C D) gp130 and (E) ICOS expression by IL-10 + (Thy1.1 + ) and IL-10 - (Thy1.1 - ) CD4 + CD3 + T cells was assessed in 10-Bit mice and shown as representative FACS plots (C) and histogram overlays (E) with mean + SEM of 5–6 mice/group (D). Gating was determined using Thy1.1 + CD4 + CD3 + cells derived from fluorescent minus one-stained samples from mice infected for 14 days. (F G) WT (C57BL/6) mice were infected with MCMV and at d6 and d10 pi αICOS or Isotype antibody was added. At d14 pi (F) salivary glands and (G) spleen CD4 + /IL-10 + responses were quantified and expressed as mean + SEM of 6 mice/group.

Techniques Used: Mouse Assay, Infection, FACS, Expressing, Derivative Assay, Staining

IL-27 promotes CD4 + IL-10 + development and impairs anti-MCMV T H 1 immunity and control of MCMV persistence. Il-27r α -/- or WT (C57BL/6) mice were infected with MCMV and spleen and salivary gland CD4 + /IFNγ + (A) responses were quantified and expressed as mean + SEM of 11 mice/group. (B) Replicating virus in salivary gland homogenates of Il-27r α -/- and WT mice is shown as individual mice + median. Data is representative of 2 experiments. (C) MCMV genomes in saliva were quantified by qPCR. Data is shown as mean ± SEM from 11 mice per group from 2 replicative experiments.

Figure Legend Snippet: IL-27 promotes CD4 + IL-10 + development and impairs anti-MCMV T H 1 immunity and control of MCMV persistence. Il-27r α -/- or WT (C57BL/6) mice were infected with MCMV and spleen and salivary gland CD4 + /IFNγ + (A) responses were quantified and expressed as mean + SEM of 11 mice/group. (B) Replicating virus in salivary gland homogenates of Il-27r α -/- and WT mice is shown as individual mice + median. Data is representative of 2 experiments. (C) MCMV genomes in saliva were quantified by qPCR. Data is shown as mean ± SEM from 11 mice per group from 2 replicative experiments.

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

6) Product Images from "Multiple sweet receptors and transduction pathways revealed in knockout mice by temperature dependence and gurmarin sensitivity"

Article Title: Multiple sweet receptors and transduction pathways revealed in knockout mice by temperature dependence and gurmarin sensitivity

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

doi: 10.1152/ajpregu.91018.2008

A : sample recordings of the integrated responses of the CT nerve of WT (C57BL/6) mice to 0.5 M sucrose (Suc) before and after treatment with gurmarin (Gur, 30 μg/ml) at three different temperatures (15, 25, and 35°C). B : Concentration-response

Figure Legend Snippet: A : sample recordings of the integrated responses of the CT nerve of WT (C57BL/6) mice to 0.5 M sucrose (Suc) before and after treatment with gurmarin (Gur, 30 μg/ml) at three different temperatures (15, 25, and 35°C). B : Concentration-response

Techniques Used: Mouse Assay, Concentration Assay

Chorda tympani (CT) nerve responses of C57BL/6 mice to 0.5 M sucrose (○), 0.02 M QHCl (▵), 0.1 M NaCl (□), 0.01 M HCl (×), 0.1 M monosodium glutamate (MSG; ◊) at different temperatures (10–45°C).

Figure Legend Snippet: Chorda tympani (CT) nerve responses of C57BL/6 mice to 0.5 M sucrose (○), 0.02 M QHCl (▵), 0.1 M NaCl (□), 0.01 M HCl (×), 0.1 M monosodium glutamate (MSG; ◊) at different temperatures (10–45°C).

Techniques Used: Mouse Assay

7) Product Images from "Hsp72 Overexpression Prevents Early Postoperative Memory Decline after Orthopedic Surgery Under General Anesthesia in Mice"

Article Title: Hsp72 Overexpression Prevents Early Postoperative Memory Decline after Orthopedic Surgery Under General Anesthesia in Mice

Journal: Anesthesiology

doi: 10.1097/ALN.0b013e31820ad3ce

Acquisition of memory during the training phase of the fear conditioning protocol is similar in C57BL/6-WT (n = 55) and Hsp72-Tg mice (n = 45). Both mouse strains achieved maximal performance by the 6 th training session. No differences in establishment

Figure Legend Snippet: Acquisition of memory during the training phase of the fear conditioning protocol is similar in C57BL/6-WT (n = 55) and Hsp72-Tg mice (n = 45). Both mouse strains achieved maximal performance by the 6 th training session. No differences in establishment

Techniques Used: Mouse Assay

8) Product Images from "The Effect of Loss of O-antigen Ligase on Phagocytic Susceptibility of Motile and Non-Motile Pseudomonas aeruginosa"

Article Title: The Effect of Loss of O-antigen Ligase on Phagocytic Susceptibility of Motile and Non-Motile Pseudomonas aeruginosa

Journal: Molecular immunology

doi: 10.1016/j.molimm.2017.10.015

Phagocytic susceptibility of O-antigen -deficient P. aeruginosa is similar to that of bacteria with O-antigen Murine bone marrow-derived dendritic cells (BMDCs) from C57BL/6 mice were assayed for relative in vitro phagocytosis of (A and C) wt PA14, motABmotCD , or waaL and (B and D) wt PAK, motABmotCD , or waaL bacteria by gentamicin protection assay incubated for 45 minutes (A and B) or 20 minutes (C and D) at MOI =10. Phagocytic uptake levels were normalized as percentages of respective mean wt phagocytosis. (E) Murine BMDCs were coincubated with wt PA14, motABmotCD, or waaL bacteria, and then fixed and probed for phospho-Akt. Akt activation (phospho-Akt) was quantified by FACS analysis and mean fluorescence intensity (MFI) was normalized to untreated cells (= 1 on Y -axis). All data are analyzed using one-way ANOVA with Tukey’s post hoc analyses and are representative of at least two independent biological experiments ( n ≥ 4). ***, p ≤ 0.0005; *, p ≤ 0.05; ns, not significant.

Figure Legend Snippet: Phagocytic susceptibility of O-antigen -deficient P. aeruginosa is similar to that of bacteria with O-antigen Murine bone marrow-derived dendritic cells (BMDCs) from C57BL/6 mice were assayed for relative in vitro phagocytosis of (A and C) wt PA14, motABmotCD , or waaL and (B and D) wt PAK, motABmotCD , or waaL bacteria by gentamicin protection assay incubated for 45 minutes (A and B) or 20 minutes (C and D) at MOI =10. Phagocytic uptake levels were normalized as percentages of respective mean wt phagocytosis. (E) Murine BMDCs were coincubated with wt PA14, motABmotCD, or waaL bacteria, and then fixed and probed for phospho-Akt. Akt activation (phospho-Akt) was quantified by FACS analysis and mean fluorescence intensity (MFI) was normalized to untreated cells (= 1 on Y -axis). All data are analyzed using one-way ANOVA with Tukey’s post hoc analyses and are representative of at least two independent biological experiments ( n ≥ 4). ***, p ≤ 0.0005; *, p ≤ 0.05; ns, not significant.

Techniques Used: Derivative Assay, Mouse Assay, In Vitro, Incubation, Activation Assay, FACS, Fluorescence

9) Product Images from "Differential ASC requirements reveal a key role for neutrophils and a noncanonical IL-1β response to Pseudomonas aeruginosa"

Article Title: Differential ASC requirements reveal a key role for neutrophils and a noncanonical IL-1β response to Pseudomonas aeruginosa

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

doi: 10.1152/ajplung.00228.2015

Neutrophils are the predominant peritoneal cellular subset that express pro-IL-1β + at early time points following P. aeruginosa infection. C57BL/6 ( n = 15) and ASC −/− ( n = 15) mice were infected intraperitoneally with a sublethal dose (3 × 10 6 CFU) of WT PA14. Mice were euthanized 4 h postinfection and peritoneal lavage was collected. A : representative dot plots depicting the gating scheme for pro-IL-1β + cells at 4 h postinfection. Bacteria were excluded by gating. B : percentage of pro-IL-1β + Ly6G + ( left ) and pro-IL-1β + F4/80 + ( right ) cells out of the total peritoneal cells at 4 h postinfection. C : percentage of Ly6G + F4/80 − ( left ) and Ly6G − F4/80 + ( right ) within the gated CD45 + pro-IL-1β + population as depicted in A above. Data are expressed as means ± SD from 5 independent biological experiments.

Figure Legend Snippet: Neutrophils are the predominant peritoneal cellular subset that express pro-IL-1β + at early time points following P. aeruginosa infection. C57BL/6 ( n = 15) and ASC −/− ( n = 15) mice were infected intraperitoneally with a sublethal dose (3 × 10 6 CFU) of WT PA14. Mice were euthanized 4 h postinfection and peritoneal lavage was collected. A : representative dot plots depicting the gating scheme for pro-IL-1β + cells at 4 h postinfection. Bacteria were excluded by gating. B : percentage of pro-IL-1β + Ly6G + ( left ) and pro-IL-1β + F4/80 + ( right ) cells out of the total peritoneal cells at 4 h postinfection. C : percentage of Ly6G + F4/80 − ( left ) and Ly6G − F4/80 + ( right ) within the gated CD45 + pro-IL-1β + population as depicted in A above. Data are expressed as means ± SD from 5 independent biological experiments.

Techniques Used: Infection, Mouse Assay

ASC −/− mice are not protected from P. aeruginosa lethal challenge and produce comparable levels of elicited in vivo IL-1β. A : C57BL/6 and ASC −/− ( n = 7/group) were infected intratracheally with 5 × 10 6 colony-forming units (CFU) of WT PA14 and survival was monitored up to 96 h postinfection. Survival curves were compared using the log-rank test. B : C57BL/6 and ASC −/− ( n = 7/group) mice were infected intratracheally with a sublethal dose (3 × 10 6 CFU) of WT PA14. C57BL/6 mice instilled with PBS ( n = 5) were used as a control. Mice were euthanized 4 h postinfection and bronchoalveolar lavage (BAL) samples were collected. BAL samples were analyzed by ELISA for IL-1β production. C : C57BL/6 ( n = 14) and ASC −/− ( n = 9) mice were infected intraperitoneally with 5 × 10 6 CFU of WT PA14 and survival was monitored up to 96 h postinfection. C57BL/6 mice were challenged with 5 × 10 6 CFU of popB mutant PA14 ( n = 8) as a control. Survival curves were compared using the log-rank test. D : C57BL/6 and ASC −/− mice were infected intraperitoneally (IP) with a sublethal dose (3 × 10 6 CFU) of WT PA14. Mice were euthanized 4 h postinfection and peritoneal lavage and blood samples were collected. Peritoneal lavage ( top ; n = 15/group) and blood serum ( bottom ; n = 11/group) samples were analyzed by ELISA for IL-1β production. Data are expressed as means ± SD. E : representative Western blot for pro- and the biologically active cleaved mature IL-1β released in the peritoneum at 4 h postinfection. Results are representative of, or accumulated from, 2–5 independent experiments.

Figure Legend Snippet: ASC −/− mice are not protected from P. aeruginosa lethal challenge and produce comparable levels of elicited in vivo IL-1β. A : C57BL/6 and ASC −/− ( n = 7/group) were infected intratracheally with 5 × 10 6 colony-forming units (CFU) of WT PA14 and survival was monitored up to 96 h postinfection. Survival curves were compared using the log-rank test. B : C57BL/6 and ASC −/− ( n = 7/group) mice were infected intratracheally with a sublethal dose (3 × 10 6 CFU) of WT PA14. C57BL/6 mice instilled with PBS ( n = 5) were used as a control. Mice were euthanized 4 h postinfection and bronchoalveolar lavage (BAL) samples were collected. BAL samples were analyzed by ELISA for IL-1β production. C : C57BL/6 ( n = 14) and ASC −/− ( n = 9) mice were infected intraperitoneally with 5 × 10 6 CFU of WT PA14 and survival was monitored up to 96 h postinfection. C57BL/6 mice were challenged with 5 × 10 6 CFU of popB mutant PA14 ( n = 8) as a control. Survival curves were compared using the log-rank test. D : C57BL/6 and ASC −/− mice were infected intraperitoneally (IP) with a sublethal dose (3 × 10 6 CFU) of WT PA14. Mice were euthanized 4 h postinfection and peritoneal lavage and blood samples were collected. Peritoneal lavage ( top ; n = 15/group) and blood serum ( bottom ; n = 11/group) samples were analyzed by ELISA for IL-1β production. Data are expressed as means ± SD. E : representative Western blot for pro- and the biologically active cleaved mature IL-1β released in the peritoneum at 4 h postinfection. Results are representative of, or accumulated from, 2–5 independent experiments.

Techniques Used: Mouse Assay, In Vivo, Infection, Enzyme-linked Immunosorbent Assay, Mutagenesis, Western Blot

Neutropenia leads to reduced peritoneal IL-1β production in response to P. aeruginosa . C57BL/6 and ASC −/− mice were depleted of neutrophils with anti-Ly6G IgG or treated with isotype control antibody ( A , C , and D ). A : representative analyses of efficacy of neutrophil depletion in circulating blood and spleens from the treatment groups. B : C57BL/6 mice were infected intraperitoneally with the indicated CFU of WT PA14. IL-1β content within the peritoneal lavage samples were subsequently assayed by ELISA( n = 2/group). C and D : C57BL/6 and ASC −/− mice were treated with isotype control antibody or depleted of neutrophils with anti-Ly6G antibody as described above. Mice were subsequently infected intraperitoneally with 3 × 10 6 CFU of WT PA14, euthanized 4 h postinfection, and peritoneal lavage and blood samples were collected for IL-1β analysis by ELISA. The peritoneal lavage fluid was analyzed for the total number of recovered CFU normalized to CFU of input bacteria. C : IL-1β from peritoneal lavage ( left ), blood serum ( middle ; BL/6, n = 10; ASC −/− , n = 7) samples and recovered peritoneal CFU from isotype IgG treated mice ( right ; n = 7/group). D : IL-1β from peritoneal lavage ( left ), blood serum ( right ; n = 10/group) and recovered peritoneal CFU from anti-Ly6G treated mice ( right ; n = 7/group). Data in A , C , and D are expressed as means ± SD and are derived from at least 3 independent biological experiments. ** P ≤ 0.05.

Figure Legend Snippet: Neutropenia leads to reduced peritoneal IL-1β production in response to P. aeruginosa . C57BL/6 and ASC −/− mice were depleted of neutrophils with anti-Ly6G IgG or treated with isotype control antibody ( A , C , and D ). A : representative analyses of efficacy of neutrophil depletion in circulating blood and spleens from the treatment groups. B : C57BL/6 mice were infected intraperitoneally with the indicated CFU of WT PA14. IL-1β content within the peritoneal lavage samples were subsequently assayed by ELISA( n = 2/group). C and D : C57BL/6 and ASC −/− mice were treated with isotype control antibody or depleted of neutrophils with anti-Ly6G antibody as described above. Mice were subsequently infected intraperitoneally with 3 × 10 6 CFU of WT PA14, euthanized 4 h postinfection, and peritoneal lavage and blood samples were collected for IL-1β analysis by ELISA. The peritoneal lavage fluid was analyzed for the total number of recovered CFU normalized to CFU of input bacteria. C : IL-1β from peritoneal lavage ( left ), blood serum ( middle ; BL/6, n = 10; ASC −/− , n = 7) samples and recovered peritoneal CFU from isotype IgG treated mice ( right ; n = 7/group). D : IL-1β from peritoneal lavage ( left ), blood serum ( right ; n = 10/group) and recovered peritoneal CFU from anti-Ly6G treated mice ( right ; n = 7/group). Data in A , C , and D are expressed as means ± SD and are derived from at least 3 independent biological experiments. ** P ≤ 0.05.

Techniques Used: Mouse Assay, Infection, Enzyme-linked Immunosorbent Assay, Derivative Assay

ASC expression is required for IL-1β production by macrophages and dendritic cells in response to Pseudomonas aeruginosa infection. Peritoneal macrophages ( A ) or bone marrow-derived dendritic cells (BMDCs; B ) from C57BL/6 or ASC −/− mice were infected with PA14 [wild-type (WT)] or with the popB mutant of PA14 at the indicated multiplicity of infection (MOI). Culture supernatants were collected 3 h postinfection and analyzed by ELISA for IL-1β production. Data are expressed as means ± SD accumulated from 2 independent biological experiments ( n = 4 for all samples except uninfected; n = 3). * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.005.

Figure Legend Snippet: ASC expression is required for IL-1β production by macrophages and dendritic cells in response to Pseudomonas aeruginosa infection. Peritoneal macrophages ( A ) or bone marrow-derived dendritic cells (BMDCs; B ) from C57BL/6 or ASC −/− mice were infected with PA14 [wild-type (WT)] or with the popB mutant of PA14 at the indicated multiplicity of infection (MOI). Culture supernatants were collected 3 h postinfection and analyzed by ELISA for IL-1β production. Data are expressed as means ± SD accumulated from 2 independent biological experiments ( n = 4 for all samples except uninfected; n = 3). * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.005.

Techniques Used: Expressing, Infection, Derivative Assay, Mouse Assay, Mutagenesis, Enzyme-linked Immunosorbent Assay

Macrophages and neutrophils temporally recruited to the peritoneum produce pro-IL-1β upon ex vivo infection. C57BL/6 mice were injected with thioglycollate and peritoneal cells were collected by lavage at 0 h (naïve uninjected), 4 h, or 3 days. The harvested cells were infected with WT or popB PA14 at MOI = 0.2 and analyzed 3 h postinfection. Bacteria were excluded from analyses by gating based on scatter controls, and the percentage of pro-IL-1β + F4/80 + and pro-IL-1β + Ly6G + cells was determined within the populations of peritoneal cells. A : representative dot plot for pro-IL-1β + cells at 0 or 4 h. B–D : percentage of pro-IL-1β + F4/80 + and pro-IL-1β + Ly6G + peritoneal cells from 0 h ( B ), 4 h ( C ), or 3 days ( D ) upon ex vivo infection. Data are expressed as means ± SD. Results are representative A , or compiled from independent biological experiments ( A–D ; n = 3).

Figure Legend Snippet: Macrophages and neutrophils temporally recruited to the peritoneum produce pro-IL-1β upon ex vivo infection. C57BL/6 mice were injected with thioglycollate and peritoneal cells were collected by lavage at 0 h (naïve uninjected), 4 h, or 3 days. The harvested cells were infected with WT or popB PA14 at MOI = 0.2 and analyzed 3 h postinfection. Bacteria were excluded from analyses by gating based on scatter controls, and the percentage of pro-IL-1β + F4/80 + and pro-IL-1β + Ly6G + cells was determined within the populations of peritoneal cells. A : representative dot plot for pro-IL-1β + cells at 0 or 4 h. B–D : percentage of pro-IL-1β + F4/80 + and pro-IL-1β + Ly6G + peritoneal cells from 0 h ( B ), 4 h ( C ), or 3 days ( D ) upon ex vivo infection. Data are expressed as means ± SD. Results are representative A , or compiled from independent biological experiments ( A–D ; n = 3).

Techniques Used: Ex Vivo, Infection, Mouse Assay, Injection

ASC is not required for pulmonary IL-1β production in response to an acute PA14 infection. C57BL/6 ( n = 7) and ASC −/− ( n = 7) mice were infected intratracheally with a sublethal dose (3 × 10 6 CFU) of WT PA14. C57BL/6 mice instilled with PBS ( n = 5) were used as a control. Mice were euthanized 4 h postinfection and BAL samples were collected. A : representative dot plot quantifying Ly6G + and CD11c + cells within a CD45 + pro-IL-1β + gate that excludes bacteria at 4 h postinfection. B : percentage of pro-IL-1β + Ly6G + ( left ) and pro-IL-1β + CD11c + ( right ) cells out of the total BAL cells at 4 h postinfection. C : percentage of Ly6G + CD11c − ( left ) and Ly6G − CD11c + ( right ) within the gated CD45 + pro-IL-1β + population as depicted in A . Data are expressed as means ± SD from 4 independent biological experiments.

Figure Legend Snippet: ASC is not required for pulmonary IL-1β production in response to an acute PA14 infection. C57BL/6 ( n = 7) and ASC −/− ( n = 7) mice were infected intratracheally with a sublethal dose (3 × 10 6 CFU) of WT PA14. C57BL/6 mice instilled with PBS ( n = 5) were used as a control. Mice were euthanized 4 h postinfection and BAL samples were collected. A : representative dot plot quantifying Ly6G + and CD11c + cells within a CD45 + pro-IL-1β + gate that excludes bacteria at 4 h postinfection. B : percentage of pro-IL-1β + Ly6G + ( left ) and pro-IL-1β + CD11c + ( right ) cells out of the total BAL cells at 4 h postinfection. C : percentage of Ly6G + CD11c − ( left ) and Ly6G − CD11c + ( right ) within the gated CD45 + pro-IL-1β + population as depicted in A . Data are expressed as means ± SD from 4 independent biological experiments.

Techniques Used: Infection, Mouse Assay

Caspase-1 dependence for neutrophil secretion of IL-1β in response to P. aeruginosa . Neutrophils were enriched by negative selection, treated with either 20 μM Z-VAD or 20 μM DCIC where indicated, and infected with WT PA14, popB PA14 or Salmonella Typhimurium at the indicated MOI. Culture supernatants were collected 3 h postinfection and analyzed by ELISA for IL-1β production ( A , B , D , E , G , and H ). A : C57BL/6 neutrophils were infected with WT PA14 or popB PA14 at a MOI = 1 in the presence or absence of Z-VAD. B : C57BL/6 and caspase-1 −/− neutrophils were infected with WT PA14, popB PA14, or S . Typhimurium at the indicated MOI and IL-1β was analyzed by ELISA. Inset : WT C57BL/6 ( lanes 1 and 2 ) or caspase-1 −/− ( lanes 3 and 4 ) neutrophils were infected with popB PA14 at a MOI = 2 ( lanes 1 and 3 ) or uninfected ( lanes 2 and 4 ), and cells were collected at 5 h postinfection and lysates analyzed for pro-IL-1β by Western analyses. C : C57BL/6 and caspase-1 −/− mice were infected intraperitoneally with a sublethal dose (3 × 10 6 CFU) of WT PA14. Mice were euthanized 4 h postinfection and peritoneal lavage and blood samples were collected. Peritoneal lavage ( top ) and blood serum ( middle ) samples were analyzed by ELISA for IL-1β production ( n ≥ 9). Pro-IL-1β was assayed from whole cell lysates of recruited peritoneal cells at 4 h postinfection ( bottom ). D : murine neutrophils or macrophages of the indicated genotype were infected with PA14 at a MOI of 1 in the presence or absence of DCIC. Data for neutrophils were compared using one-way ANOVA with the Tukey-Kramer posttest. E : C57BL/6 and caspase-1 −/− neutrophils were infected with WT PA14 at MOI = 1 in the presence or absence of DCIC or Z-VAD. Data were compared using one-way ANOVA with the Tukey-Kramer posttest. F : C57BL/6 neutrophils were infected with popB mutant PA14 at MOI = 2 in the presence or absence of Z-VAD or DCIC, and subsequently analyzed by Western blot with the same protocol used in B , inset . G : C57BL/6 and neutrophil elastase −/− neutrophils were infected with WT PA14 or popB mutant PA14 at the indicated MOI. H : C57BL/6 and neutrophil elastase −/− neutrophils were infected with WT PA14 at MOI = 1 in the presence or absence of Z-VAD. Data were compared using one-way ANOVA with the Tukey-Kramer posttest. Results are representative of 2 ( A , n = 4; D , n = 6; E , n = 6; F and G , n = 6; H , n = 6) and 3 independent biological experiments ( B , n = 6; C , n ≥ 9). Data are expressed as means ± SD. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.005.

Figure Legend Snippet: Caspase-1 dependence for neutrophil secretion of IL-1β in response to P. aeruginosa . Neutrophils were enriched by negative selection, treated with either 20 μM Z-VAD or 20 μM DCIC where indicated, and infected with WT PA14, popB PA14 or Salmonella Typhimurium at the indicated MOI. Culture supernatants were collected 3 h postinfection and analyzed by ELISA for IL-1β production ( A , B , D , E , G , and H ). A : C57BL/6 neutrophils were infected with WT PA14 or popB PA14 at a MOI = 1 in the presence or absence of Z-VAD. B : C57BL/6 and caspase-1 −/− neutrophils were infected with WT PA14, popB PA14, or S . Typhimurium at the indicated MOI and IL-1β was analyzed by ELISA. Inset : WT C57BL/6 ( lanes 1 and 2 ) or caspase-1 −/− ( lanes 3 and 4 ) neutrophils were infected with popB PA14 at a MOI = 2 ( lanes 1 and 3 ) or uninfected ( lanes 2 and 4 ), and cells were collected at 5 h postinfection and lysates analyzed for pro-IL-1β by Western analyses. C : C57BL/6 and caspase-1 −/− mice were infected intraperitoneally with a sublethal dose (3 × 10 6 CFU) of WT PA14. Mice were euthanized 4 h postinfection and peritoneal lavage and blood samples were collected. Peritoneal lavage ( top ) and blood serum ( middle ) samples were analyzed by ELISA for IL-1β production ( n ≥ 9). Pro-IL-1β was assayed from whole cell lysates of recruited peritoneal cells at 4 h postinfection ( bottom ). D : murine neutrophils or macrophages of the indicated genotype were infected with PA14 at a MOI of 1 in the presence or absence of DCIC. Data for neutrophils were compared using one-way ANOVA with the Tukey-Kramer posttest. E : C57BL/6 and caspase-1 −/− neutrophils were infected with WT PA14 at MOI = 1 in the presence or absence of DCIC or Z-VAD. Data were compared using one-way ANOVA with the Tukey-Kramer posttest. F : C57BL/6 neutrophils were infected with popB mutant PA14 at MOI = 2 in the presence or absence of Z-VAD or DCIC, and subsequently analyzed by Western blot with the same protocol used in B , inset . G : C57BL/6 and neutrophil elastase −/− neutrophils were infected with WT PA14 or popB mutant PA14 at the indicated MOI. H : C57BL/6 and neutrophil elastase −/− neutrophils were infected with WT PA14 at MOI = 1 in the presence or absence of Z-VAD. Data were compared using one-way ANOVA with the Tukey-Kramer posttest. Results are representative of 2 ( A , n = 4; D , n = 6; E , n = 6; F and G , n = 6; H , n = 6) and 3 independent biological experiments ( B , n = 6; C , n ≥ 9). Data are expressed as means ± SD. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.005.

Techniques Used: Selection, Infection, Enzyme-linked Immunosorbent Assay, Western Blot, Mouse Assay, Mutagenesis

10) Product Images from "The Tripeptide KdPT Protects from Intestinal Inflammation and Maintains Intestinal Barrier Function"

Article Title: The Tripeptide KdPT Protects from Intestinal Inflammation and Maintains Intestinal Barrier Function

Journal: The American Journal of Pathology

doi: 10.1016/j.ajpath.2011.05.013

Effect of KdPT at various concentrations after i.p. administration and after oral and rectal application in DSS-induced colitis. C57BL/6 mice received various amounts of KdPT by daily i.p. injections starting 2 days after induction of DSS colitis. Inflammation

Figure Legend Snippet: Effect of KdPT at various concentrations after i.p. administration and after oral and rectal application in DSS-induced colitis. C57BL/6 mice received various amounts of KdPT by daily i.p. injections starting 2 days after induction of DSS colitis. Inflammation

Techniques Used: Mouse Assay

KdPT ameliorates DSS-induced colitis. C57BL/6 WT mice received 3% DSS in their drinking water for 5 days, and inflammation was monitored by daily measurement of individual weights. A: From day 2 onward, one group was treated with 10 μg of KdPT

Figure Legend Snippet: KdPT ameliorates DSS-induced colitis. C57BL/6 WT mice received 3% DSS in their drinking water for 5 days, and inflammation was monitored by daily measurement of individual weights. A: From day 2 onward, one group was treated with 10 μg of KdPT

Techniques Used: Mouse Assay

11) Product Images from "Maturational differences in lung NF-?B activation and their role in tolerance to hyperoxia"

Article Title: Maturational differences in lung NF-?B activation and their role in tolerance to hyperoxia

Journal: Journal of Clinical Investigation

doi: 10.1172/JCI200419300

Demonstration of hyperoxia-mediated neonatal (Neo) lung NF-κB activation in C57BL/6 mice. ( A ) Representative of 3 EMSA blots. F, free probe; 0–72 h, duration of hyperoxia; C, competition with 100-fold excess of unlabeled oligonucleotide probe; M, competition with 100-fold excess of unlabeled mutated oligonucleotide probe; S, supershift with p50 antibody in lung extract collected at 8 hours of hyperoxia; SS, supershift retardation band. ( B ) Densitometric evaluation of the EMSA blots. Densitometric units were expressed as a ratio of air-exposed values. The data are the mean ± SE of 3 separate experiments. * P

Figure Legend Snippet: Demonstration of hyperoxia-mediated neonatal (Neo) lung NF-κB activation in C57BL/6 mice. ( A ) Representative of 3 EMSA blots. F, free probe; 0–72 h, duration of hyperoxia; C, competition with 100-fold excess of unlabeled oligonucleotide probe; M, competition with 100-fold excess of unlabeled mutated oligonucleotide probe; S, supershift with p50 antibody in lung extract collected at 8 hours of hyperoxia; SS, supershift retardation band. ( B ) Densitometric evaluation of the EMSA blots. Densitometric units were expressed as a ratio of air-exposed values. The data are the mean ± SE of 3 separate experiments. * P

Techniques Used: Activation Assay, Mouse Assay

12) Product Images from "The H2B deubiquitinase Usp22 promotes antibody class switch recombination by facilitating non-homologous end joining"

Article Title: The H2B deubiquitinase Usp22 promotes antibody class switch recombination by facilitating non-homologous end joining

Journal: Nature Communications

doi: 10.1038/s41467-018-03455-x

IgG1 CSR primarily relies on c-NHEJ, whereas IgA CSR is more dependent on A-EJ. a Spleen B cells isolated from 53BP1 KO or WT littermates were induced for ex vivo IgG1, IgG2b, IgG3, or IgA CSR for 4 days. 53BP1 −/− B cells (deficient in c-NHEJ) exhibited a less severe defect in IgA CSR (~ 60% reduction) compared with IgG CSR (~ 80–90% reduction). Data represent two independent experiments each with three to four mice per group. b A-196 (10 µM) was used to inhibit c-NHEJ during the ex vivo CSR of WT spleen B cells and A-197 is the control compound. Inhibition of c-NHEJ markedly reduced IgG1 CSR, but not IgA CSR ( n = 4 samples per group). Data represent three independent experiments. c , d The switch region sequence of Sμ on the IgH locus of C57BL/6 background mice were compared with that of c Sα or d Sγ1. Dot Matrix analysis of the mouse switch regions. The dots represent homologies between two switch regions with a search length of 30 bp and defined percentage of identity. Data in a and b were presented as mean ± SEM and were analyzed using two-tailed unpaired Student’s t -test. *** p

Figure Legend Snippet: IgG1 CSR primarily relies on c-NHEJ, whereas IgA CSR is more dependent on A-EJ. a Spleen B cells isolated from 53BP1 KO or WT littermates were induced for ex vivo IgG1, IgG2b, IgG3, or IgA CSR for 4 days. 53BP1 −/− B cells (deficient in c-NHEJ) exhibited a less severe defect in IgA CSR (~ 60% reduction) compared with IgG CSR (~ 80–90% reduction). Data represent two independent experiments each with three to four mice per group. b A-196 (10 µM) was used to inhibit c-NHEJ during the ex vivo CSR of WT spleen B cells and A-197 is the control compound. Inhibition of c-NHEJ markedly reduced IgG1 CSR, but not IgA CSR ( n = 4 samples per group). Data represent three independent experiments. c , d The switch region sequence of Sμ on the IgH locus of C57BL/6 background mice were compared with that of c Sα or d Sγ1. Dot Matrix analysis of the mouse switch regions. The dots represent homologies between two switch regions with a search length of 30 bp and defined percentage of identity. Data in a and b were presented as mean ± SEM and were analyzed using two-tailed unpaired Student’s t -test. *** p

Techniques Used: Non-Homologous End Joining, Isolation, Ex Vivo, Mouse Assay, Inhibition, Sequencing, Two Tailed Test

13) Product Images from "Type I Interferon Production Induced by Streptococcus pyogenes-Derived Nucleic Acids Is Required for Host Protection"

Article Title: Type I Interferon Production Induced by Streptococcus pyogenes-Derived Nucleic Acids Is Required for Host Protection

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1001345

Type I IFN signaling is needed for host defense against S. pyogenes and control of neutrophil recruitment. ( A ) Kaplan-Meier survival curves of C57BL/6 and IFNAR1 -/- mice (15 mice per genotype) after subcutaneous infection with 3×10 8 CFU of S. pyogenes . Survival was monitored for 6 days. Significance: ** = P value

Figure Legend Snippet: Type I IFN signaling is needed for host defense against S. pyogenes and control of neutrophil recruitment. ( A ) Kaplan-Meier survival curves of C57BL/6 and IFNAR1 -/- mice (15 mice per genotype) after subcutaneous infection with 3×10 8 CFU of S. pyogenes . Survival was monitored for 6 days. Significance: ** = P value

Techniques Used: Mouse Assay, Infection

14) Product Images from "Beneficial effects of IL-37 after spinal cord injury in mice"

Article Title: Beneficial effects of IL-37 after spinal cord injury in mice

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

doi: 10.1073/pnas.1523212113

Infusion of rIL-37 promotes functional recovery after SCI. Locomotor performance of C57BL/6 mice treated with intraspinal injections of saline, full-length IL-37b (IL-37 1–218 ), or the processed form of IL-37b (IL-37 V46–218 ) after SCI using

Figure Legend Snippet: Infusion of rIL-37 promotes functional recovery after SCI. Locomotor performance of C57BL/6 mice treated with intraspinal injections of saline, full-length IL-37b (IL-37 1–218 ), or the processed form of IL-37b (IL-37 V46–218 ) after SCI using

Techniques Used: Functional Assay, Mouse Assay

15) Product Images from "MyD88 Signaling in B Cells Regulates the Production of Th1-dependent Antibodies to AAV"

Article Title: MyD88 Signaling in B Cells Regulates the Production of Th1-dependent Antibodies to AAV

Journal: Molecular Therapy

doi: 10.1038/mt.2012.101

Humoral response to adeno-associated virus serotype 1 (AAV1) vector is TLR/IL-1R and type I interferon (IFN)-R-independent but MyD88-dependent. Control C57Bl/6 or mutant TLR9 −/− , TLR7 −/− , TRIF −/− , 3D, TLR2 −/− , TLR4 −/− , MyD88 −/− , IFNαβR −/− , caspase 1 −/− , NALP3 −/− , IL-1R1 −/− mice were infused intravenous (i.v.) with 10 11 vg of rAAV1-U7mEx23 vector. Serum samples were harvested at weeks 4–5 for the measurement of anti-rAAV1 immunoglobulin G (IgG) titers. Histograms represent the geometric mean ± SEM of reciprocal end-point titers of 2 to 4 independent experiments for each targeted mutant ( n = 6–13 mice per group).

Figure Legend Snippet: Humoral response to adeno-associated virus serotype 1 (AAV1) vector is TLR/IL-1R and type I interferon (IFN)-R-independent but MyD88-dependent. Control C57Bl/6 or mutant TLR9 −/− , TLR7 −/− , TRIF −/− , 3D, TLR2 −/− , TLR4 −/− , MyD88 −/− , IFNαβR −/− , caspase 1 −/− , NALP3 −/− , IL-1R1 −/− mice were infused intravenous (i.v.) with 10 11 vg of rAAV1-U7mEx23 vector. Serum samples were harvested at weeks 4–5 for the measurement of anti-rAAV1 immunoglobulin G (IgG) titers. Histograms represent the geometric mean ± SEM of reciprocal end-point titers of 2 to 4 independent experiments for each targeted mutant ( n = 6–13 mice per group).

Techniques Used: Plasmid Preparation, Mutagenesis, Mouse Assay

16) Product Images from "Role of CCR8 and Other Chemokine Pathways in the Migration of Monocyte-derived Dendritic Cells to Lymph Nodes"

Article Title: Role of CCR8 and Other Chemokine Pathways in the Migration of Monocyte-derived Dendritic Cells to Lymph Nodes

Journal: The Journal of Experimental Medicine

doi: 10.1084/jem.20032152

Allostimulatory capacity of monocyte subsets sorted from the periphery and effect of an antagonist to the CCR8 ligand CCL1. (A) Gr-1 hi and Gr-1 int recruited monocytes from the peritoneal lavage of C57BL/6 CCR8 +/+ mice were sorted to purity by flow cytometry using mAbs to F4/80 and Gr-1. The cells were cultured separately in GM-CSF for 2 d in the absence of added mAb, or in the presence of 5 μg/ml of neutralizing mAbs to chemokines CCL1 (TCA-3) or CCL19. These cells were washed to remove residual antibodies and cultured with BALB/c T cells for evaluation of their potential to support allogeneic T cell proliferation in a mixed lymphocyte reaction. These data depict one out of two experiments conducted with similar results.

Figure Legend Snippet: Allostimulatory capacity of monocyte subsets sorted from the periphery and effect of an antagonist to the CCR8 ligand CCL1. (A) Gr-1 hi and Gr-1 int recruited monocytes from the peritoneal lavage of C57BL/6 CCR8 +/+ mice were sorted to purity by flow cytometry using mAbs to F4/80 and Gr-1. The cells were cultured separately in GM-CSF for 2 d in the absence of added mAb, or in the presence of 5 μg/ml of neutralizing mAbs to chemokines CCL1 (TCA-3) or CCL19. These cells were washed to remove residual antibodies and cultured with BALB/c T cells for evaluation of their potential to support allogeneic T cell proliferation in a mixed lymphocyte reaction. These data depict one out of two experiments conducted with similar results.

Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Cell Culture

Analysis of cell-mediated microsphere transport to LNs in CCR8 −/− and plt/plt mice. (A) Green fluorescent LX microspheres were injected into the skin of CCR8-deficient mice and plt/plt mice that were compared with age- and sex-matched WT C57BL/6 counterparts. The number of DCs bearing two or more LX particles in the draining LNs was quantified 3 d later. To combine data from different experiments, the mean number of migrated cells in WT mice was set equal to 1.0 for each experiment, and relative values for all WT and knock-out individuals in that experiment were calculated. Inset shows Gr-1 staining intensity among gated bead + cells from CCR8 −/− (bold line) and CCR8 +/+ mice (thin line) in the skin 14 h after bead injection. (B and C) Plots show MHC II (I-A b ) and Gr-1 levels in LNs and skin of WT and CCR8 −/− mice. LN plots (B) are quantitative comparisons, as they depict the entire population of LX-bearing cells recovered from pooled brachial LNs from individual mice. Skin plots (C) are not quantitative comparisons. Where available, plots depict all acquired events, although in some experiments (for MHC II staining in LN), LX + cells were gated during acquisition to reduce file size. (D) High power magnification of WT skin section stained for LYVE-1 (green) and CCL1 (red). (E) LYVE-1 (green) and CCL1 (red) in LN subcapsular sinus (left, high power magnification). CCL1 staining in LN at low power (right). Outer subcapsule is indicated by an arrow.

Figure Legend Snippet: Analysis of cell-mediated microsphere transport to LNs in CCR8 −/− and plt/plt mice. (A) Green fluorescent LX microspheres were injected into the skin of CCR8-deficient mice and plt/plt mice that were compared with age- and sex-matched WT C57BL/6 counterparts. The number of DCs bearing two or more LX particles in the draining LNs was quantified 3 d later. To combine data from different experiments, the mean number of migrated cells in WT mice was set equal to 1.0 for each experiment, and relative values for all WT and knock-out individuals in that experiment were calculated. Inset shows Gr-1 staining intensity among gated bead + cells from CCR8 −/− (bold line) and CCR8 +/+ mice (thin line) in the skin 14 h after bead injection. (B and C) Plots show MHC II (I-A b ) and Gr-1 levels in LNs and skin of WT and CCR8 −/− mice. LN plots (B) are quantitative comparisons, as they depict the entire population of LX-bearing cells recovered from pooled brachial LNs from individual mice. Skin plots (C) are not quantitative comparisons. Where available, plots depict all acquired events, although in some experiments (for MHC II staining in LN), LX + cells were gated during acquisition to reduce file size. (D) High power magnification of WT skin section stained for LYVE-1 (green) and CCL1 (red). (E) LYVE-1 (green) and CCL1 (red) in LN subcapsular sinus (left, high power magnification). CCL1 staining in LN at low power (right). Outer subcapsule is indicated by an arrow.

Techniques Used: Mouse Assay, Injection, Knock-Out, Staining

Mutagenesis:

Article Title: Essential role for histone deacetylase 11 (HDAC11) in neutrophil biology
Article Snippet: .. C57BL/6 WT mice were purchased from Charles River Laboratories (Wilmington, MA, USA), Tg‐HDAC11‐eGFP reporter mice [ ] were provided by the Nathaniel Heintz Institute through the Mutant Mouse Resource & Research Centers, and HDAC11KO, kindly supplied by Merck (Kenilworth, NJ, USA) and obtained from the E.S. lab, respectively. .. All strains of mice were housed in the same designated room at the animal facility (Vincent A. Stabile Research Building, Moffitt Cancer Center, Tampa, FL, USA), kept in a pathogen‐free condition, and handled in accordance with the requirements of the U.S.

Cell Culture:

Article Title: The Effect of Loss of O-antigen Ligase on Phagocytic Susceptibility of Motile and Non-Motile Pseudomonas aeruginosa
Article Snippet: .. Bone marrow-derived dendritic cells (BMDCs) were cultured from C57BL/6 WT mice (Charles River Laboratories) as previously described [ ]. ..

Mouse Assay:

Article Title: Cytomegalovirus-Specific IL-10-Producing CD4+ T Cells Are Governed by Type-I IFN-Induced IL-27 and Promote Virus Persistence
Article Snippet: .. C57BL/6 WT mice were purchased from Charles River or Envigo. .. Mice were infected with MCMV that was prepared by sorbital gradient purification as described previously [ ].

Article Title: Dendritic cell functional improvement in a preclinical model of lentiviral-mediated gene therapy for Wiskott-Aldrich syndrome
Article Snippet: .. C57BL/6 wt mice and OVA specific, MHC class II restricted, TCR transgenic OT-II mice were purchased from Charles River Laboratories Inc. (Calco, Italy). .. These mice were housed under specific pathogen-free (SPF) conditions and treated according to protocols approved by the Animal Care and Use Committee of the San Raffaele Scientific Institute (IACUC 318).

Article Title: Multiple sweet receptors and transduction pathways revealed in knockout mice by temperature dependence and gurmarin sensitivity
Article Snippet: .. C57BL/6 WT mice were obtained from the Charles River Japan (Tokyo, Japan). ..

Article Title: Both TRIF and IPS-1 Adaptor Proteins Contribute to the Cerebral Innate Immune Response against Herpes Simplex Virus 1 Infection
Article Snippet: .. C57BL/6 WT mice were purchased from Charles River Canada (St-Constant, Quebec, Canada). .. TRIF−/− and IPS-1−/− mice (both provided by S. Akira, Osaka University, Osaka, Japan) were generated and maintained in a C57BL/6 background as described previously ( , ).

Article Title: Hsp72 Overexpression Prevents Early Postoperative Memory Decline after Orthopedic Surgery Under General Anesthesia in Mice
Article Snippet: .. C57BL/6-WT mice were purchased from Charles River Laboratories (Wilmington, MA). .. Adult male mice were randomly allocated to one of three treatment groups: control (naïve animals without treatment; (C), anesthesia alone (A) and tibial fracture surgery (S).

Article Title: Conditional Expression of Human 15-Lipoxygenase-1 in Mouse Prostate Induces Prostatic Intraepithelial Neoplasia: The FLiMP Mouse Model 1
Article Snippet: .. All three transgenic founder lines were bred to C57BL/6 wt mice (NCI Charles River Laboratories) and were characterized for germline transmission, tissue tropism, and level of CAT expression. .. In addition, multiple tissues from all lines were tested for basal h15-LO-1 expression to ensure that there was no aberrant background expression of 15-LO-1.

c57bl 6 wt mice (Charles River Laboratories)

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