Journal:

Article Title: TraJ-Dependent Escherichia coli K1 Interactions with Professional Phagocytes Are Important for Early Systemic Dissemination of Infection in the Neonatal Rat

doi: 10.1128/IAI.72.1.478-488.2004

Figure Lengend Snippet: E. coli K1 associates with professional phagocytes during early dissemination. Representative immunohistology analysis of neonatal rat spleens 1 day post-oral infection with E. coli K1 is shown. Tissue samples were subjected to immunofluorescent staining for dendritic cell marker Ox-62 and the E. coli K1 wild type (A and C) and the traJ mutant (B) via step-wise permeabilization as described in Materials and Methods. (A and B) Intracellular bacteria were stained with Cy5.5 (aqua), and extracellular bacteria were stained with Cy3 (red). Tissue sections were also stained with dendritic-cell-specific cell marker Ox-62 (green) and the cell nuclear material stain DAPI (blue). Stained sections were then examined, and images were captured with a Leica DM-RXA fluorescence microscope. (C) Intracellular bacteria were stained with Cy3 (red), and extracellular bacteria were stained with Cy5.5 (aqua). Stained sections were then examined with a Leica SP1 confocal microscope, and images were displayed as xy, xz, and yz orthogonal projections.

Article Snippet: Ten-micrometer sections were subjected to immunofluorescent microscopy with the following antibodies: anti- E. coli K1 (Abcam, Inc.), anti-ED-2 (Research Diagnostics, Inc.), anti-Ox-62 (Research Diagnostics, Inc.), anti-ED-1 (Research Diagnostics, Inc.), anti-mouse immunoglobulin G-fluorescein isothiocyanate (IgG-FITC) (Sigma-Aldrich), anti-mouse IgG-Cy3 (Sigma-Aldrich), and anti-mouse IgG-Cy5.5 (Rockland, Inc.).

Techniques: Infection, Staining, Marker, Mutagenesis, Fluorescence, Microscopy

Journal: Genes & Development

Article Title: Interaction of CCR4–NOT with EBF1 regulates gene-specific transcription and mRNA stability in B lymphopoiesis

doi: 10.1101/gad.285452.116

Figure Lengend Snippet: Cnot3 -deficient mice show impaired early B-cell differentiation that can be alleviated in part by the expression of Mzb1 . ( A ) Flow cytometric analysis of bone marrow B cells in Cnot3 +/+ RERT Cre and Cnot3 fl/fl RERT Cre mice to detect B220 + CD43 + pro-B, B220 int CD43 − late pre-B, and immature B cells as well as B220 hi CD43 − recirculating B cells in the living lymphocyte gate. ( B ) Further gating of pro-B cells to detect pre-pro-B (HSA − BP1 − ), early pro-B (HSA + BP1 − ) and late pro-B (HSA + BP1 + ) cells. ( C ) Statistical analysis of the total numbers of pro-B (pink) and pre-B (red) cells in Cnot3 +/+ RERT Cre (wild-type) and Cnot3 fl/fl RERT Cre (knockout) mice. Long horizontal bars indicate the mean, and short horizontal bars represent the SD of the mean. Statistical significance between wild type and knockout is assessed by an unpaired two-tail Student's t -test. ( D ) PCR analysis of genomic DNA from Cnot3 +/+ RERT Cre and Cnot3 fl/fl RERT Cre mice to detect the deletion status of Cnot3 . Cnot6 served as a loading control. ( E ) Flow cytometric analysis of splenic B cells to detect CD21 int CD23 + follicular B cells and CD21 hi CD23 − marginal zone B cells in Cnot3 +/+ RERT Cre and Cnot3 fl/fl RERT Cre mice. ( F ) Flow cytometric analysis of bone marrow cells in Cnot3 +/+ mb1 Cre and Cnot3 fl/fl mb1 Cre mice to detect the B-cell compartment as described in A . ( G ) Flow cytometric analysis of the effects of a Mzb1 transgene expression ( Mzb1 tg ) on early B-cell differentiation in Cnot3 +/+ mb1 Cre and Cnot3 fl/fl mb1 Cre mice. ( H , I ) Gating of bone marrow B220 + CD43 + pro-B cells to detect HSA + BP1 − early pro-B and HSA + BP1 + late pro-B stages. ( J , K ) Statistical analysis of the total numbers of pre-B (red), recirculating B (blue), early pro-B (orange), and late pro-B (green) cells in Cnot3 +/+ mb1 Cre , Cnot3 fl/fl mb1 Cre , Cnot3 +l+ mb1 Cre Mzb1 tg , and Cnot3 fl/fl mb1 Cre Mzb1 tg mice. Numbers in the FACS profiles indicate the percentage of cells within the gated population. The data are representative of four or more independent experiments.

Article Snippet: Anti-CD19 (1D3), BP1 (BP-1), CD25 (PC61) HSA (M1/69), CD43 (R2/60), B220 (RA-6B2), CD93 (AA4.1), CD23 (B3B4), CD21 (7G6), and α-IgMμ (Jackson ImmunoResearch, #115–175-075) were used.

Techniques: Cell Differentiation, Expressing, Knock-Out, Control