Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: SC transplantation shifted the CD11b immune cell population from an Arg1 − iNOS + pro-inflammatory to an intermediate Arg1 + iNOS + phenotype after SCI. Representative images of flow cytometry analysis and pie charts of CD11b population dynamics at 14 days post-injury (7 days post-transplantation) show, compared with SCI controls ( A – C ), a decreased percentage of CD11b cells stained with Arg1 − iNOS + and an increased percentage for Arg1 + iNOS + in animals receiving SC transplants ( D – F ). Results are expressed as mean ± standard deviation (SD). Abbreviations on the graphs are: Fluorescein isothiocyanate (FITC), Allophycocyanin (APC) and Forward Scatter (FSC-A). For panels ( B , E ), the blue dots represent the CD11b population that is iNOS − -Arg1 − , the orange dots represent the CD11b population that is iNOS + -Arg1 − and the green dots represent the CD11b population that is double positive for both iNOS + -Arg1 + . These colored dots are also shown in the forward scatter plots of panels ( A , D ).

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Transplantation Assay, Flow Cytometry, Staining, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: CD68 immune cells with a pro-inflammatory Arg1 − iNOS + phenotype after SCI are reduced by SC transplantation. Representative images of flow cytometry analysis and pie charts of CD68 population dynamics at 14 days post-injury (7 days post-transplantation) reveal that CD68 immune cells with a pro-inflammatory Arg1 − iNOS + phenotype after SCI ( A – C ) are reduced by the intraspinal transplantation of SCs ( D – F ). Results are expressed as mean ± SD. For panels ( A , D ), the orange dots represent the ED1 population that were gated based on their forward and side scatter from the total events (blue) that were acquired. The orange dots in the different quadrants of ( B , E ) represent the ED1 population that was positive for either Arg1 (top left quadrant) or iNOS (bottom right quadrant) or double positive for both markers (top right quadrant).

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Transplantation Assay, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: CD11b immune cells expressing a highly pro-inflammatory CD38 + iNOS + phenotype or anti-inflammatory Arg1 + CD163 + form after SCI were unaltered by SC transplantation. Representative images of flow cytometry analysis and pie charts of CD11b population dynamics at 14 days post-injury (7 days post-transplantation) reveal that CD11b immune cells with a highly pro-inflammatory CD38 + iNOS + phenotype after SCI ( A , B ) are not altered by SC transplantation ( E , F ). Similarly, CD11b immune cells with a highly anti-inflammatory Arg1 + CD163 + phenotype were unchanged across SCI controls ( C , D ) and SC-transplanted groups ( G , H ). Results are expressed as mean ± SD. For panels ( A , B ), the blue dots represent the CD11b population that were CD38 − iNOS − , the orange dots represent the CD11b population that were CD38 − iNOS + , the gray dots represent the CD11b population that were CD38 + iNOS − and the green dots represent the CD11b population that were double positive for both CD38 + iNOS + . For panels ( C , D ), the configuration for the colored dots is the same for the representation labeling, single, double or absent, though the proteins Arg1 and CD163 are represented rather than CD38 and iNOS.

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Expressing, Transplantation Assay, Flow Cytometry, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: Cytokine profile of CD11b + cells is unaltered by SC implants after SCI. Representative images of flow cytometry analysis and pie charts of CD11b population dynamics at 14 days post-injury (7 days post-transplantation) show that CD11b immune cells have unaltered expression of pro- (TNF-α, IL-1β) and anti-inflammatory (IL-4, IL-10) cytokines comparatively among SCI controls ( A – E ) and SC-transplanted groups ( F – J ). Results are expressed as mean ± SD. For panels ( A – D , F – I ) the blue dots represent the cell population that were CD11b − or did not express the selected cytokines, whereas the green dots represent the CD11b + population that expressed CD11b and the cytokine of interest (top right quadrant).

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Flow Cytometry, Transplantation Assay, Expressing

Journal: bioRxiv

Article Title: Lymph node resident memory T cells retain effector capabilities by evading lung resident memory dysfunction

doi: 10.64898/2026.02.03.703569

Figure Lengend Snippet: (A) Naive CD90.1+ OT-I T cells were transferred into naive B6 mice before intranasal infection with influenza expressing SIINFEKL (PR8-OVA). Expression of CD69 and CD103 was measured 21 days post infection on extravascular cells. (B) Quantification of CD69 + CD103 + OT-I T cells. (C) Naïve CD90.1 + OT-I Nur77-GFP T cells were transferred into mice one day before intranasal PR8-OVA infection. Nur77-GFP expression was evaluated 3-14 days later by extravascular OT-I T cells. (D)Experimental setup for (E-G). Flow cytometry plot (E) of CD90.1 + OT-I T cells isolated from indicated locations. (F-G) Longitudinal labeling of cells and quantification. (H) Experimental setup for (I-J). (I) Labeling after intravenous and intraperitoneal injection of OT-I T cells or naïve endogenous T cells. (J) Expression of CD69 and CD103. Naïve= CD8 + CD90.1 - CD44 low . Each point represents a mouse and N=2-5 mice per experiment. Data are representative (A) or are pooled from at least two experiments (n=2–4/group), except for day 113 in (B), which reflect a single experiment.

Article Snippet: Measurements were obtained in Flow Cytometry (FACS) Staining Buffer (Rockland, MB-086-0500) at room temperature for about 30 minutes per sample and channels were flushed with buffer in between samples.

Techniques: Infection, Expressing, Flow Cytometry, Isolation, Labeling, Injection

Journal: bioRxiv

Article Title: Lymph node resident memory T cells retain effector capabilities by evading lung resident memory dysfunction

doi: 10.64898/2026.02.03.703569

Figure Lengend Snippet: (A)Chromatin accessibility with gene expression (right) and linked chromatin peaks (bottom) at the Gzmb locus. (B) Protein expression of GranzymeB as measured by flow cytometry. (C-D) Same as (A-B) but for the Pdcd1 locus. Protein expression on OT-I T cells is 49 days post infection where each dot represents one mouse. (J and L) Data are from one experiment and representative of at least two experiments with N=3. (E-F) Surface expression of PD-1 and TIM3 at 7 days post infection. (G) Transcription factor motif footprint analysis from dataset. (H) Inferred eRegulons identified by SCENIC+ across memory T cells after influenza infection. (I-K) Legend (left) for network plots illustrating inferred transcription factor relationships to target genes. Transcription factor and gene relationship is denoted by color of line represents activation (red) or repression (blue).

Article Snippet: Measurements were obtained in Flow Cytometry (FACS) Staining Buffer (Rockland, MB-086-0500) at room temperature for about 30 minutes per sample and channels were flushed with buffer in between samples.

Techniques: Gene Expression, Expressing, Flow Cytometry, Infection, Activation Assay

Journal: International Journal of Molecular Sciences

Article Title: Molecular Characterization of Gastric Epithelial Cells Using Flow Cytometry

doi: 10.3390/ijms19041096

Figure Lengend Snippet: Gating strategy for analyzing gastric epithelial cells by flow cytometry. ( A ) A gate based of forward area and side scatter area is first established, these cells are then put through a forward scatter area and forward scatter height gate to identify single cells, finally, 7-AAD is used to separate live cells from dead/dying cells; ( B ) Representative flow plots of live single cells from healthy BALB/c and TxA23 mice stained with an epithelial cell marker (EpCAM) and an immune cell marker (CD45). Immune cells are undetectable in the gastric mucosa control mice, and present in TxA23 mice that have autoimmune gastritis; ( C ) Representative immunocytochemistry of glands isolated from a 2 month old BALB/c mouse and stained with hoechst (blue), parietal cell marker Dolichous bifluorous agglutinin (DBA, green), and anti-EpCAM (red) with a high magnification inset in yellow showing an individual gland; ( D ) A representative flow cytometry plot of live single cells from a BALB/c mouse stained with anti-EpCAM and DBA demonstrating a subset of EpCAM + DBA + parietal cells.

Article Snippet: Following Dispase digestion, single cells were washed twice using flow cytometry staining buffer supplemented with 20 mM EDTA (Promega, Madison, WI, USA, V4231) and counted using Trypan blue exclusion.

Techniques: Flow Cytometry, Staining, Marker, Control, Immunocytochemistry, Isolation

Journal:

Article Title: Gamma Interferon Is Not Required for Arthritis Resistance in the Murine Lyme Disease Model

doi: 10.1128/IAI.69.6.3737-3743.2001

Figure Lengend Snippet: Flow cytometry of CD4+ T cells. CD4+ T cells producing IL-4 predominated in the IFN-γR−/− mice, while those producing IFN-γ predominated in the 129/SvEv mice at all time points tested. Mice were sacrificed 10 days postinfection and LN cells restimulated in vitro with B. burgdorferi antigen-pulsed irradiated syngeneic spleen cells. At the indicated times cells were stained for CD4, IL-4, and IFN-γ and analyzed by flow cytometry. (A) Log10 fluorescence of blast cells on day 8 is presented for a representative experiment of three; (B) Percent blast cells with the Th2 (CD4+ IL-4+) or Th1 (CD4+ IFN-γ+) phenotype for 129/SvEv and IFN-γR−/− mice, calculated from results of flow cytometry, as a function of time in vitro.

Article Snippet: Flow cytometry staining buffer I (FSB-I) consisted of PBS (BioWhittaker) with 1% heat-inactivated fetal calf serum (Sigma Chemical Co.) and 0.1% NaN 3 (Sigma Chemical Co.), and FSB-II was PBS with 1% heat-inactivated fetal calf serum and 0.1% saponin (Sigma Chemical Co.).

Techniques: Flow Cytometry, In Vitro, Irradiation, Staining, Fluorescence

Journal:

Article Title: Gamma Interferon Is Not Required for Arthritis Resistance in the Murine Lyme Disease Model

doi: 10.1128/IAI.69.6.3737-3743.2001

Figure Lengend Snippet: Flow cytometry of CD8+ T cells and macrophages. Levels of CD8+ cells producing IFN-γ and macrophages producing IL-12 were higher in the 129/SvEv than IFN-γR−/− mice. Restimulated cultures of mock-infected mice had few cells, and cultures from 129/SvEv and IFN-γR−/− mice generated approximately equivalent numbers of IL-12+ MAC-1+ cells (0.7% and 0.5%, respectively). Results presented are for culture day 8 and are from an experiment representative of three.

Article Snippet: Flow cytometry staining buffer I (FSB-I) consisted of PBS (BioWhittaker) with 1% heat-inactivated fetal calf serum (Sigma Chemical Co.) and 0.1% NaN 3 (Sigma Chemical Co.), and FSB-II was PBS with 1% heat-inactivated fetal calf serum and 0.1% saponin (Sigma Chemical Co.).

Techniques: Flow Cytometry, Infection, Generated

Journal: International Journal of Biological Sciences

Article Title: N -Acetylcysteine overcomes epalrestat-mediated increase of toxic 4-hydroxy-2-nonenal and potentiates the anti-arthritic effect of epalrestat in AIA model

doi: 10.7150/ijbs.85028

Figure Lengend Snippet: Epalrestat enhances autoimmune response in AIA rats. Blood lymphocytes were harvested from epalrestat-treated AIA rats for flow cytometry analysis using fluorescent antibodies against CD45, CD3, CD4, CD8 and IL-17A. (a) Representative flow cytometry images show the percentage of CD3 + T lymphocytes gated on CD45 + lymphocytes. The bar chart shows the proportion of CD3 + T cells among CD45 + lymphocytes. (b) Representative flow cytometry images show the proportion of CD4 + T cells and CD8 + T cells gated on CD3 + T lymphocytes. The quantitative bar chart shows the ratio of CD4 + / CD8 + T cells. (c) Representative flow cytometry images show the percentage of the IL-17A + T cells gated on CD4 + T lymphocytes. The bar chart shows the percentage of IL-17A + T cells among CD4 + T cells. (d) The serum concentration of inflammatory cytokines in epalrestat-treated AIA rats. The concentration of these cytokines was measured using LEGENDplex TM Rat Inflammatory Panel. The data are presented as the means ± SEM (n ≥5). *P<0.05, **P<0.01, ***P<0.001, significant different compared with healthy control or vehicle control.

Article Snippet: Cells were then washed and resuspended in 200 cL flow cytometry staining buffer and analyzed using FACSAria III cell sorter (BD Biosciences, USA).

Techniques: Flow Cytometry, Concentration Assay

Journal: International Journal of Biological Sciences

Article Title: N -Acetylcysteine overcomes epalrestat-mediated increase of toxic 4-hydroxy-2-nonenal and potentiates the anti-arthritic effect of epalrestat in AIA model

doi: 10.7150/ijbs.85028

Figure Lengend Snippet: NAC reverses the epalrestat-induced autoimmune response in AIA rats. Blood lymphocytes were harvested from epalrestat and NAC combined treated AIA rats for flow cytometry analysis using fluorescent antibodies against CD45, CD3, CD4, CD8, IL-17A and Foxp3. (a) Representative flow cytometry images show the percentage of CD4 + and CD8 + T lymphocytes gated on CD3 + T lymphocytes, and the percentage of the Foxp3 + T cell or IL-17A + T cells gated on CD4 + T lymphocytes. (b) The quantitative bar charts show the ratio of CD4 + / CD8 + T cells, and the percentage of IL-17A + T cells or Foxp3 + T cells among CD4 + T cells. (c&d) Co-treatment of NAC reversed the changed serum concentration of cytokines in AIA rats. The concentration of different cytokines was measured using LEGENDplex TM Customer Panel. The data are presented as the means ± SEM (n ≥5). *P<0.05, **P<0.01, ***P<0.001, compared with healthy control or vehicle control.

Article Snippet: Cells were then washed and resuspended in 200 cL flow cytometry staining buffer and analyzed using FACSAria III cell sorter (BD Biosciences, USA).

Techniques: Flow Cytometry, Concentration Assay