Journal: Frontiers in Microbiology

Article Title: A Functional Slow Recycling Pathway of Transferrin is Required for Growth of Chlamydia

doi: 10.3389/fmicb.2010.00112

Figure Lengend Snippet: Analysis of host cell trafficking pathways during treatment with FR179254 . HEp-2 cells were treated as indicated, and trafficking pathways were analyzed as described in Section (A) Lipid droplet (LD) visualization with the neutral lipid stain Bodipy 493. (B) Fluid-phase uptake as measured by fluorescent dextran. (C) Live cell visualization of multivesicular bodies/late endosomes using CD63-GFP expressing HEp-2 cells. (D) Analysis of late endosomes/lysosomes in LAMP1-YFP transfected cells. (E) Exocytic trafficking as measured by incorporation of fluorescent sphingomyelin (SM) in Golgi and chlamydiae. (F) Exocytic trafficking of glycoproteins as measured by localization of CD4 on the surface of transfected cells. (G,H) Analysis of retrograde trafficking using fluorescently tagged cholera toxin subunit B (CTxB) after pulse-chase labeling. Arrows within (C) and (E) indicate inclusions. Note the lack of significant staining in (C) (inset) and the presence of staining in (E) , which is contrast to a published study (Beatty, ).

Article Snippet: For CD4, non-permeabilized cells were incubated with primary Ab mouse anti-human CD4 (R&D Systems, Minneapolis, MN, USA) followed by secondary goat anti-mouse Alexa488 (Invitrogen).

Techniques: Staining, Expressing, Transfection, Pulse Chase, Labeling

Journal: Journal of Respiration

Article Title: Immune Correlates of Non-Necrotic and Necrotic Granulomas in Pulmonary Tuberculosis: A Pilot Study

doi: 10.3390/jor1040023

Figure Lengend Snippet: Figure 2. Distribution of CD15, S100A8/A9, CD4 and IBA1 positive immune cells in human lung TB granulomas. (A) Representative images singleplex (CD15 and S100A8/A9; red spots) and multiplex (CD4; green spots and IBA1; red spots) immunostaining of control, solid/non-necrotic and necrotic granulomas. Host cell nucleus is stained blue with DAPI. Scale bar in S100A8/A9 panel (50 microns) is common for CD15 and S100A8/A9 images. Scale bar in the IBA1 panel (50 microns) is common for CD4 and IBA1 images. (B) Distribution of CD15 positive cells (including MDSCs) relative to the total number of cells in the field in control, solid/non-necrotic granulomas and necrotic granulomas. (C) Distribution of S100A8/A9 positive cells (mainly neutrophils) relative to the total number of cells in the field in control, solid/non-necrotic granulomas and necrotic granulomas. (D) Distribution of CD4 positive T-cells relative to the total number of cells in the field in control, solid/non-necrotic granulomas and necrotic granulomas. (E) Distribution of IBA1 positive cells (activated macrophages) relative to the total number of cells in the field in control, solid/non-necrotic granulomas and necrotic granulomas. Data plotted in (B–E) are mean and standard error mean (SEM) of n = 3 (control); n = 5 (solid lesion) and n = 18 (necrotic lesion). Data were analyzed by One-way ANOVA with Tukey’s post-hoc correction for multiple group comparison; * p < 0.05; ** p < 0.01; *** p < 0.005.

Article Snippet: S100A8/A9) (cat no. ab22506) were purchased from (Abcam, Waltham, MA, USA), CD33 (cat no. sc19660), CD15 (cat no. sc-19649), ARG1 and NOS2 (cat no. sc-7271) (Santa Cruz Biotechnology, Dallas, TX, USA), HIF-1α (cat no. MA5160048) Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA), IL-6 (cat no. MAB206) and IL-10 (cat no. AF-217-NA) (R& D Systems Inc, Minneapolis, MN, USA) and CD4 from (cat no. NBP1-19371AF647) Novus Biologicals, Centennial, CO, USA).

Techniques: Multiplex Assay, Immunostaining, Control, Staining, Comparison