rabbit polyclonal anti gb  (Cusabio)

Journal: Nature Communications

Article Title: Streptococcus agalactiae cadD alleviates metal stress and promotes intracellular survival in macrophages and ascending infection during pregnancy

doi: 10.1038/s41467-022-32916-7

Figure Lengend Snippet: A – C Immunofluorescence microscopy examination of the placenta of either uninfected ( A ) or GBS-infected ( B ) mice reveals macrophages within the tissue as determined by staining with a monoclonal antibody to F4/80 (red). Tissues were also stained with a polyclonal antibody to GBS (green), and counter-stained with 4′,6-diamidino-2-phenylindole (DAPI; blue) to visualize cell nuclei. Merged images reveal co-localization of GBS and macrophages (yellow), magnification bar indicates 50 μm. C Enlargement of inset panel of the merged image in B indicates GBS cells associated with tissue macrophages within the placenta (yellow arrows). Micrographs are shown which are representative of images collected from 3 separate dams from each experimental group (uninfected or GBS-infected, n = 3). D , E GBS cultured within THP-1 macrophages in vitro were subjected to transcriptional analyses. D RNAseq analysis revealed that cadD was upregulated after 24 h of culture within macrophages, a result that was confirmed by quantitative real-time PCR ( E ), bars indicate mean and error bars indicate +/− SEM. * P = 0.0083, two-tailed, paired Student’s t test, n = 3 separate biological replicates. F Gene sequencing indicates that cadD is organized within a short operon and is highly conserved across numerous Streptococcus spp . including oral streptococci. The maximum likelihood phylogeny was constructed with 500 bootstrap replications; bootstrap support values are indicated at each node, scale bar indicates number of substitutions per site.

Article Snippet: Slides were incubated with a 1:100 dilution of rabbit polyclonal anti-GBS antibody (ab78846; Abcam) for 1 h. The Bond Polymer Refine detection system (Leica Biosystems) was used for visualization.

Techniques: Immunofluorescence, Microscopy, Infection, Staining, Cell Culture, In Vitro, Real-time Polymerase Chain Reaction, Two Tailed Test, Sequencing, Construct

Journal: Nature Communications

Article Title: Streptococcus agalactiae cadD alleviates metal stress and promotes intracellular survival in macrophages and ascending infection during pregnancy

doi: 10.1038/s41467-022-32916-7

Figure Lengend Snippet: A Conceptual diagram of methods utilized in these studies. Pregnant mice were infected with GBS on embryonic day 13.5 and sacrificed 2 days post-infection. Reproductive tissues were collected for analyses. B Bacterial burden within reproductive tissues was evaluated by quantitative culture to determine Log CFU/mg (vaginal tissue was collected from 5 dams per experimental group, uterus tissues were collected from 6 separate dams, and all other tissues were collected from 6 separate fetal-placental units per experimental group). Significant changes in burden were observed between the WT GB112 and ∆cadD mutant in the decidua (* P = 0.0257), placenta (* P = 0.0003), amnion (* P = 0.0312), and fetus (* P = 0.0266) by one-tailed Mann–Whitney U analysis. These results were reversed by genetic complementation in trans resulting in significant changes in burden observed between the ∆cadD mutant and the complemented mutant in the decidua (* P = 0.0185), placenta (* P = 0.0285), amnion (* P = 0.0009), and fetus (* P = 0.0006) by one-tailed Mann–Whitney U analysis. C Histopathological examination of the hematoxylin and eosin-stained placental units at ×100 magnification (inset panel at ×400 magnification). D Placental units were analyzed by immunohistochemical techniques using a polyclonal antibody to stain specifically for GBS (indicated by the brown stain). Microscopic imaging of reproductive tissues from pregnant mice infected with WT GBS (GB112), isogenic ΔcadD mutant ( ΔcadD ), and the complemented isogenic ΔcadD mutant ( ΔcadD:C ). Micrographs are representative of analyses performed in a blinded fashion on tissues derived from 6 separate dams per experimental group. Magnification bars indicate 250 μm. All statistical analyses performed were one-tailed analyses.

Article Snippet: Slides were incubated with a 1:100 dilution of rabbit polyclonal anti-GBS antibody (ab78846; Abcam) for 1 h. The Bond Polymer Refine detection system (Leica Biosystems) was used for visualization.

Techniques: Infection, Mutagenesis, One-tailed Test, MANN-WHITNEY, Staining, Immunohistochemical staining, Imaging, Derivative Assay

Journal: Nature Communications

Article Title: Streptococcus agalactiae cadD alleviates metal stress and promotes intracellular survival in macrophages and ascending infection during pregnancy

doi: 10.1038/s41467-022-32916-7

Figure Lengend Snippet: A Conceptual diagram of methods utilized in these studies. Pregnant mice were intraperitoneally injected on embryonic day 12.5 and 14.5 with either anti-F4/80 antibody to deplete host macrophages, or an isotype control antibody. Mice were infected with GBS on embryonic day 13.5 and sacrificed 2 days post-infection. Vaginal tissues were collected from three separate dams for each experimental condition for analyses. Uterine tissues were collected from four separate dams for each experimental condition for analyses. For decidual, placental, amnion, and fetal tissues, each individual data point indicates an independent fetal-placental unit. B Flow cytometry confirms that macrophages were depleted within the decidua and placenta at the maternal-fetal interface. C Placental units were analyzed by immunohistochemical techniques using a polyclonal antibody to stain specifically for GBS (indicated by the brown stain). Micrographs are representative of three biological replicates which were analyzed in a blinded fashion. Microscopical imaging at ×100 magnification reveals bacterial invasion of the reproductive tract in pregnant animals infected with WT GBS (GB112), the ΔcadD mutant ( ΔcadD ), and the complemented isogenic ΔcadD mutant ( ΔcadD:C ). Micrographs were analyzed in a blinded fashion and are representative of three biological replicates. D Bacterial burden within reproductive tissues was evaluated by quantitative culture (Log CFU/mg) in response to macrophage depletion via anti-F4/80 injections (+) compared to isotype control treatments (−) lacking macrophage depletion. Significant changes in burden were observed in WT GB112-infected mice between the isotype control and the anti-F4/80 treatment groups in the decidua (* P = 0.020), placenta (* P = 0.0008), amnion (* P = 0.0312), and fetus (* P = 0.0266) by one-tailed Mann–Whitney U analysis. These results were also observed in the cohort of animals infected with the complemented mutant in the decidua (* P = 0.0385), placenta (* P = 0.0130), amnion (* P = 0.0011), and fetus (* P = 0.0242) by one-tailed Mann–Whitney U test, but not in the animals infected with the isogenic ∆cadD mutant (NS = not significant).

Article Snippet: Slides were incubated with a 1:100 dilution of rabbit polyclonal anti-GBS antibody (ab78846; Abcam) for 1 h. The Bond Polymer Refine detection system (Leica Biosystems) was used for visualization.

Techniques: Injection, Control, Infection, Flow Cytometry, Immunohistochemical staining, Staining, Imaging, Mutagenesis, One-tailed Test, MANN-WHITNEY

Journal: Aging (Albany NY)

Article Title: Dietary type 2 resistant starch improves systemic inflammation and intestinal permeability by modulating microbiota and metabolites in aged mice on high-fat diet

doi: 10.18632/aging.103187

Figure Lengend Snippet: RS2 reduced intestinal permeability and inflammation in blood, colon, and liver in aged mice on high-fat diet. ( A ) Representative images of PAS-AB staining in colon tissues from three groups (400x magnification, scale bar =50 μm) and the quantification of PAS-AB positive cells per crypt for every mouse in each group. ( B ) Immunofluorescence staining of MUC2 (green) with nuclear counterstaining (blue) in colon tissue from the three groups (200x magnification, scale bar=100 μm) and quantification analysis of mean fluorescence intensity for every mouse in each group. ( C ) Effects of diet on colonic MUC2 mRNA expression assessed by quantitative real-time PCR. ( D ) Comparison of serum LPS levels assessed by ELISA among the three groups. ( E ) Comparison of fecal LPS levels assessed by ELISA among the three groups. ( F ) Effects of diets on colonic and hepatic inflammatory cytokines mRNA expression assessed by quantitative real-time PCR. Multiple comparisons of colonic TNF-α levels (p=0.22); multiple comparisons of hepatic TNF-α levels (p=0.07). n=5 or 6/group. Data are expressed as mean+SE. Differences were compared by one-way ANOVA among the three groups with Tukey’s multiple comparison posttests between two groups. * p<0.05, ** p<0.01, *** p<0.001. MUC2, mucin2; CON, control group; HF, high-fat diet group; HFRS, high-fat diet+20%RS2 group.

Article Snippet: Sections were blocked for 30 mins at room temperature in bovine serum albumin (BSA) to prevent nonspecific staining and incubated with primary antibody polyclonal rabbit anti-MUC2 antibody (1:800, Servicebio, GB 11344, Wuhan, China) overnight at 4°C.

Techniques: Permeability, Staining, Immunofluorescence, Fluorescence, Expressing, Real-time Polymerase Chain Reaction, Comparison, Enzyme-linked Immunosorbent Assay, Control

Journal: Aging (Albany NY)

Article Title: Dietary type 2 resistant starch improves systemic inflammation and intestinal permeability by modulating microbiota and metabolites in aged mice on high-fat diet

doi: 10.18632/aging.103187

Figure Lengend Snippet: Primer sequences.

Article Snippet: Sections were blocked for 30 mins at room temperature in bovine serum albumin (BSA) to prevent nonspecific staining and incubated with primary antibody polyclonal rabbit anti-MUC2 antibody (1:800, Servicebio, GB 11344, Wuhan, China) overnight at 4°C.

Techniques: