Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: Validation of small extracellular vesicles. ( A ) The workflow of the isolation of small extracellular vesicles (sEVs) from the plasma of healthy controls (HCs) and recovered COVID-19 patients (RCs), including recovered asymptomatic patients (RAs), recovered moderate patients (RMs), and recovered severe and critical patients (RSs). ( B ) Representative transmission electron microscopy of the morphology of sEVs using commercial Exosome kit from RAs, RMs, RSs, and HCs. Scale bar: 200 nm. ( C ) Nanoparticle tracking analysis of sEVs from the plasma of RAs, RMs, RSs, and HCs. The detailed data were presented in Tables and . ( D ) The size distribution of the purified sEVs isolated from plasma of HCs, RAs, RMs, and RSs. ( E ) Western blotting indicates the expression levels of CD9, CD63, TSG101, Calnexin, GM130, GRP94, and Lamin B1 in cells, RAs, RMs, RSs, and HCs. Among them, cells were selected as control. The gels were cropped from different parts and automatically exposed using BIORAD. Cropped blots were displayed alongside their corresponding original blots in Fig. .

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques: Biomarker Discovery, Isolation, Clinical Proteomics, Transmission Assay, Electron Microscopy, Purification, Western Blot, Expressing, Control

Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: ( A ) Percentage of identified sEVs cargo lipid classes and subclasses. ( B ) Relative abundance (%) of sEVs lipids subclasses in recovered asymptomatic patients (RAs), recovered moderate patients (RMs), recovered severe/critical patients (RSs), and healthy controls (HCs). ( C – E ) The level of total lipids and lipids subclasses diacylglycerols (DAGs) and phosphatidylcholines (PCs) in RAs, RMs, RSs, and HCs. Data were displayed as boxplots with median and range and each dot represents an individual: HC (Pink), RA (Green), RM (Red), RS (Blue). p-value: *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques:

Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: Lipid profiles of sEVs isolated from HCs, RAs, RMs, and RSs plasma. ( A – C ) Orthogonal partial least square discriminant analysis score plot in recovered asymptomatic patients (RAs), recovered moderate patients (RMs), and recovered severe/critical patients (RSs) compared to HCs. ( D ) The heatmap shows significantly altered lipids in the sEVs isolated from the plasma of RAs, RMs, RSs, and HCs. Only differential lipids with p < 0.05 were displayed, and the color shades indicated the levels of lipids. Yellow and blue are indicative of relatively higher and lower levels, respectively.

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques: Isolation, Clinical Proteomics

Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: ( A – C ) Volcano plots show the sEVs differential lipids in RAs ( A ), RMs ( B ), or RSs ( C ) compared with HCs. The x-axis was the value of log2(FC), FC indicates the ratio of the mean level of the sEVs lipid in RAs, RMs, or RSs to the mean value of HCs, y-axis was − lg(p-value). Blue represented downregulation with p < 0.05, red represented upregulation, and grey denoted no change. ( D ) Venn diagram shows the number of significantly altered lipids in RAs, RMs, and RSs relative to HCs. ( E ) Statistical analysis of FFA(22:5) was displayed as bar graphs in sEVs of HCs, RAs, RMs, and RSs. The significance of comparisons was determined by the Kruskal–Wallis test. Data were displayed as boxplots with median and range and each dot represents an individual: HC (Pink), RA (Green), RM (Red), RS (Blue). Lipids quantitated using targeted lipidomics were presented in nanograms of lipids per milliliter (ng/mL) sEVs. p-value: *p < 0.05; **p < 0.01; ***p < 0.001. RAs recovered asymptomatic patients, RMs recovered moderate patients, RSs recovered severe/critical patients.

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques:

Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: Distinct lipid profiles in RCs (RAs, RMs, and RSs) and KEGG enrichment analysis. ( A – C ) Volcano plots presenting differential lipids in RM vs. RA, RS vs. RA, and RS vs. RM. The x-axis is the log2(FC) value, and the y-axis is the − lg (p-value). Blue represented downregulation with p < 0.05, red represented upregulation, and grey denoted no change. ( D ) Venn plot showing the differential lipids among diverse RC groups (RAs, RMs, and RSs). The blue, red, and green circles represent RM vs. RA, RS vs. RA, and RS vs. RM groups. ( E ) Statistical analysis of DAG(16:1/18:0) is displayed as the box plot graph. The significance of comparisons was determined by the Kruskal–Wallis test. Lipids quantitated using targeted lipidomics were presented in nanomoles of lipids per milliliter (ng/mL) sEVs. p-value: *p < 0.05; **p < 0.01; ***p < 0.001. ( F ) Lipidomic KEGG enrichment analysis of differential lipids in RCs relative to HCs. The pathway enrichment analysis calculates the p-value and pathway impact, respectively. The node color is based on the p-value, and the node radius is determined based on pathway impact values.

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques:

Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: Correlation of plasma sEVs lipids with clinical indices. ( A – C ) Correlation plots illustrate Spearman correlations between clinical indices with differential lipids identified in RAs ( A ), RMs ( B ), and RSs ( C ) relative to HCs in 102 recruiters. Only correlation with p < 0.05 was indicated with colored squares and marked with stars (*). Positive and negative correlations were shown in blue and red, respectively, with sizes of squares representing the magnitude of the correlations. *p < 0.05; ** p < 0.01; ***p < 0.001. RAs recovered asymptomatic patients, RMs recovered moderate patients, RSs recovered severe/critical patients.

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques: Clinical Proteomics

Journal: Cancer Control : Journal of the Moffitt Cancer Center

Article Title: Clinical Characteristics and Prognosis of Small Cell Carcinoma in the Nasopharynx: A Population-Based Study

doi: 10.1177/10732748221087075

Figure Lengend Snippet: ( A ) Hematoxylin and eosin (H&E) staining of the nasopharyngeal tumor specimen under ×100 magnification. ( B ) Hematoxylin and eosin (H&E) staining of the nasopharyngeal tumor specimen under ×200 magnification. The tumor mass is immunopositive for ( C ) CD56, ( D ) creatine kinase (CK), ( E ) Ki67, and ( F ) synaptophysin (SYN; ×200 magnification); it is immunonegative for ( F ) chromogranin A (CgA), ( G ) EBER, ( H ) p40, and ( I ) p63.

Article Snippet: Hematoxylin and eosin (H&E) staining was performed on tumor tissue samples, followed by immunohistochemical staining with Ki67 rabbit monoclonal antibody (1:400, #9027, Cell Signaling Technology, Boston, the United States), CD56 mouse monoclonal antibody (1:800, #3576, Cell Signaling Technology, Boston, the United States), pan-keratin mouse monoclonal antibody (1:200, #67306, Cell Signaling Technology, Boston, the United States), synaptophysin (SYN) rabbit monoclonal antibody (1:200, #36406, Cell Signaling Technology, Boston, the United States), chromogranin A (CgA) rabbit monoclonal antibody (MAB-0707, MXB Biotechnologies, Fuzhou, China), p40 monoclonal antibody (RMA-0815, MXB Biotechnologies, Fuzhou, China), and p63 rabbit monoclonal antibody (1:450, #39692, Cell Signaling Technology, Boston, the United States).

Techniques: Staining

Journal: International immunopharmacology

Article Title: Compromised adenosine-A2AR axis contributes to recurrent spontaneous abortion by promoting proinflammatory macrophage polarization.

doi: 10.1016/j.intimp.2025.114838

Figure Lengend Snippet: Fig. 1. RSA patients had lower levels of CD39 and adenosine. (A) Flow cytometry gating strategy for the analysis of live+ CD45+ dNK cells in decidual tissues isolated from patients. (B) Flow cytometry staining of CD39 on dNK cells in decidual tissues from normal pregnancies (HC) (n = 12) and RSA patients (n = 10). (C) Immunofluorescence staining for CD39 (green), CD56 (red), and DAPI (blue) in decidual tissues. Yellow arrows indicate CD56+ dNK cells with high expression of CD39. (D) Quantification of CD56+ CD39+ positive cells per field (n = 4). Scale bars: 50 μm. (E) dNK cells were isolated from decidual tissues of normal pregnant women (n = 12) and RSA patients (n = 10), co-cultured with HTR-8/SVneo cells in 24-well plates (dNK: HTR-8/SVneo cells = 2:1) for 24 h and adenosine levels in the media were measured by ELISA. Data are expressed as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The cryostat sections were fixed with 4 % PFA and incubated in the blocking buffer at room temperature for 1 h. Sections were incubated overnight at 4 ◦C with primary antibodies: mouse anti-human CD39 (Abcam, cat: ab97552, dilution 1:100) and rabbit anti-human CD56 (CST, cat: 99746, dilution 1:100), followed by 2 h incubation with secondary antibodies: FITC-conjugated goat anti-mouse IgG (H + L) (ZSGB-BIO, cat: ZF0312, dilution 1:50) and Rhodamine-conjugated goat anti-rabbit IgG (ZSGB-BIO, cat: ZF0316, dilution 1:50).

Techniques: Flow Cytometry, Isolation, Staining, Immunofluorescence, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Cell death & disease

Article Title: Scutellarin activates IDH1 to exert antitumor effects in hepatocellular carcinoma progression.

doi: 10.1038/s41419-024-06625-6

Figure Lengend Snippet: Fig. 7 Scu exerts antitumor effects by promoting IDH1 enzyme activity and activating the tumor immune microenvironment in vivo. A Schematic plan for the administration of Scu (60 and 100 mg/kg/day). C The tumor weight and D tumor volume were monitored every week for four weeks. After the mice were sacrificed, the resected tumors were B photographed and processed for pathological and immunohistochemical assays for E, F necrosis area and Ki67, L, M HIF1α, GLUT1, VEGFA, and N, O CD4, CD8, F4/80, CD56 and PD-L1 expression. G The relative mRNA levels of the indicated genes, H, I the expression levels of the indicated proteins, J the level of α-KG, and K IDH1 activity in tumor tissue were detected after treatment with Scu in vivo. The black arrows indicate positively stained cells. Scale bars, 50 μm. n ≥5. The data are mean ± SD; *P < 0.05; **P < 0.01; ***P < 0.001 compared to the control group.

Article Snippet: The sections were blocked in PBS containing 10% normal goat serum and 0.3% Triton X-100 for 60min; labeled with primary antibodies against Ki67 (1:2000, 27309-1-AP, Proteintech), IDH1 (1:200, 12332-1-AP, Proteintech), HIF1a (1:200, A22041, ABclonal), GLUT1 (1:200, 81463-1-RR, Proteintech), VEGFA (1:100, A12303, ABclonal), CD4 (1:500, 67786-1-Ig, Proteintech), CD8 (1:10,000, 66868-1-Ig, Proteintech), F4/80 (1:100, A23788, ABclonal), CD56 (1:2000, 14255-1-AP, Proteintech), and PDL1 (1:1000, 28076-1-AP, Proteintech) overnight at 4 °C; and incubated with the corresponding goat secondary antibodies for 1 h at room temperature.

Techniques: Activity Assay, In Vivo, Immunohistochemical staining, Expressing, Staining, Control

Journal: PLOS Pathogens

Article Title: CD56-mediated activation of human natural killer cells is triggered by Aspergillus fumigatus galactosaminogalactan

doi: 10.1371/journal.ppat.1012315

Figure Lengend Snippet: CD56 expression on naïve NK cells (Control) and NK cells stimulated for 3 h with A . fumigatus germ tubes, depending on pre-treatment of the germ tubes with proteinase K (A) or periodate (B). Data for 3 independent donors are shown. (C) Significantly differentially expressed genes after 3-h stimulation of NK cells with A . fumigatus germ tubes pre-treated (60 min) with periodate or untreated (0 min). Abbreviations: adj. p = adjusted p-value, D = donor, fc = fold change. (D) Volcano plot summarizing transcriptional changes in NK cells stimulated with periodate-treated A . fumigatus germ tubes compared to cells stimulated with untreated germ tubes. Genes with a log2 fold change > 0.5 and an adjusted p-value < 0.05 are highlighted. Selected immune-related genes have been labelled. (E) CD56 expression on naïve NK cells (Control) and NK cells stimulated for 3 h with A . fumigatus germ tubes, depending on the duration of periodate pre-treatment of the germ tubes. Representative histograms and data for cells from 5 independent donors are shown. (F) Chemokine and perforin release by naïve NK cells (Control) and NK cells stimulated for 3 h with A . fumigatus germ tubes, depending on the duration of periodate pre-treatment of the germ tubes. (A-B; E-F) Columns and error bars indicate means and standard deviations, respectively. Repeated measures one-way analysis of variance with Tukey’s post-hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Hyphae were incubated with soluble CD56 (5 μg/mL, R&D Systems, Minneapolis, MN, USA, Cat#2408-NC) or BSA (control, 5 μg/mL, Roth, Karlsruhe, Germany, #0163.2) in colorless RPMI for 2 h at 37°C, 5% CO 2 .

Techniques: Expressing, Control

Journal: PLOS Pathogens

Article Title: CD56-mediated activation of human natural killer cells is triggered by Aspergillus fumigatus galactosaminogalactan

doi: 10.1371/journal.ppat.1012315

Figure Lengend Snippet: (A) CD56 binding to fungal carbohydrates and proteins, as determined by enzyme-linked immunosorbent assay. GAG = galactosaminogalactan, RodAp = surface rodlet protein/hydrophobin. N = 3 technical replicates. One-way analysis of variance (ANOVA) with Dunnett’s post-hoc test versus Control, i.e., no coating. (B) Pull-down assay of CD56 with urea-insoluble galactosaminogalactan (PGG) or β-1,3-glucan. Lane M: Protein marker; Lane A: loading buffer extract of GAG + CD56 pellet; Lane B: supernatant of GAG + CD56; Lane C: CD56 alone; Lane D: supernatant of β-1,3-glucan + CD56; Lane E: loading buffer extract of β-1,3-glucan + CD56 pellet. (C) CD56 and CD69 expression on naïve NK cells (Control) and NK cells stimulated for 24 h with different concentrations of PGG. Representative histograms and data for cells isolated from 3 independent donors are shown. (D) Chemokine release by naïve NK cells (Control) and NK cells stimulated for 24 h with different concentrations of PGG. N = 3 independent donors. (C-D) Repeated measures (RM) one-way ANOVA with Tukey’s post-hoc test. (E) CD56, CD69, and CD107a expression on naïve NK cells (Control) and NK cells stimulated for 6 h with A . fumigatus (ATCC46645, AF) or A . nidulans (ATCC11267, AN) at different multiplicities of infection (MOIs). Representative histograms for one donor at MOI 4 are shown. (F) MOI-dependent stimulation of NK-cellular chemokine secretion after 6-h stimulation with AF or AN. (E-F) N = 3 independent donors. RM one-way ANOVA with Dunnett’s post-hoc test versus Control, i.e., unstimulated NK cells (asterisks). In addition, AF and AN stimulation at each MOI was compared using paired t-Test (hash signs). (A-F) Columns and error bars indicate means and standard deviations, respectively. */# p < 0.05, **/## p < 0.01, *** p < 0.001.

Article Snippet: Hyphae were incubated with soluble CD56 (5 μg/mL, R&D Systems, Minneapolis, MN, USA, Cat#2408-NC) or BSA (control, 5 μg/mL, Roth, Karlsruhe, Germany, #0163.2) in colorless RPMI for 2 h at 37°C, 5% CO 2 .

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Control, Pull Down Assay, Marker, Expressing, Isolation, Infection

Journal: PLOS Pathogens

Article Title: CD56-mediated activation of human natural killer cells is triggered by Aspergillus fumigatus galactosaminogalactan

doi: 10.1371/journal.ppat.1012315

Figure Lengend Snippet: (A) Representative fluorescent micrographs (z-projection of 3–4 slices with 1 μm distance, representative dataset from ≥5 independent experiments) of hyphae of wild-type (WT) A . fumigatus Af293 and two galactosaminogalactan (GAG)-deficient A . fumigatus mutants (Δ uge3 and Δ agd3 ) co-cultured with soluble CD56, followed by staining with fluorescent anti-CD56 antibody. The shape of the hyphae is indicated by dotted lines The insets represent a control image (bottom; BSA stained with anti-CD56 antibody) of the respective strain. Scale: 10 μm. (B) CD56, CD69, and CD107a expression on naïve NK cells (Control) and NK cells stimulated for 6 h with WT Af293 or the GAG-deficient A . fumigatus mutants (Δ uge3 and Δ agd3 ) at different multiplicities of infection (MOIs). Representative histograms for one donor at MOI 4 are shown. (C) MOI-dependent induction of NK-cellular secretion of granzyme B, perforin, and chemokines after 6-h stimulation with WT Af293, Δ uge3 , and Δ agd3 . (B-C) N = 3 independent donors. Repeated measures (RM) one-way analysis of variance (ANOVA) with Dunnett’s post-hoc test versus Control, i.e., unstimulated NK cells (asterisks). In addition, results for stimulation with the 3 strains at each MOI was compared using RM one-way ANOVA with Dunnett’s post-hoc test versus WT (hash signs). Columns and error bars indicate means and standard deviations, respectively. */# p < 0.05, **/## p < 0.01, ***/### p < 0.001. (D) CLSM micrographs of NK cells co-cultured with WT Af293 and Δ agd3 A . fumigatus hyphae. CD56 was stained with anti-CD56 Alexa Fluor 647 (red) to assess the CD56 localization. Germ tubes could be detected via their auto-fluorescence (cyan). Scale: 10 μm.

Article Snippet: Hyphae were incubated with soluble CD56 (5 μg/mL, R&D Systems, Minneapolis, MN, USA, Cat#2408-NC) or BSA (control, 5 μg/mL, Roth, Karlsruhe, Germany, #0163.2) in colorless RPMI for 2 h at 37°C, 5% CO 2 .

Techniques: Cell Culture, Staining, Control, Expressing, Infection, Fluorescence

Journal: PLOS Pathogens

Article Title: CD56-mediated activation of human natural killer cells is triggered by Aspergillus fumigatus galactosaminogalactan

doi: 10.1371/journal.ppat.1012315

Figure Lengend Snippet: (A) CD56 binding to fully acetylated (aPGG), fully deacetylated (dePGG), and native galactosaminogalactan (PGG), was determined by enzyme-linked immunosorbent assay (ELISA). Additional conditions with incomplete ELISA setup were included to preclude unspecific binding of the secondary antibody or anti-CD56 to the carbohydrates. N = 3 technical replicates. One-way analysis of variance (ANOVA) with Tukey’s post-hoc test was performed for each assay setup. (B) CD56, CD69, and CD107a expression on naïve NK cells (Control) and NK cells stimulated for 24 h with PGG, aPGG, or dePGG. Representative histograms for cells isolated from one donor are shown. (C) NK-cellular chemokine secretion after 24-h stimulation with PGG, aPGG, or dePGG. (B-C) N = 6 independent donors. Repeated measures (RM) one-way ANOVA with Tukey’s post-hoc test. (D) CD56, CD69, and CD107a expression on naïve NK cells (Control) and NK cells stimulated for 24 h with GalN oligomers, GalNAc oligomers, or chitosan. NK-cellular chemokine secretion after 24-h stimulation with GalN oligomers, GalNAc oligomers, or chitosan. N = 4 independent donors. RM one-way ANOVA with Tukey’s post-hoc test. (A-D) * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Hyphae were incubated with soluble CD56 (5 μg/mL, R&D Systems, Minneapolis, MN, USA, Cat#2408-NC) or BSA (control, 5 μg/mL, Roth, Karlsruhe, Germany, #0163.2) in colorless RPMI for 2 h at 37°C, 5% CO 2 .

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Expressing, Control, Isolation

Journal: PLOS Pathogens

Article Title: CD56-mediated activation of human natural killer cells is triggered by Aspergillus fumigatus galactosaminogalactan

doi: 10.1371/journal.ppat.1012315

Figure Lengend Snippet: (A) CD56 and CD69 expression of unstimulated NK cells (Control) and A . fumigatus -stimulated NK cells (6 h) depending on the fungal strain (wild type [WT] Af293 or isogenic mutants with defective GAG biosynthesis) and its enzymatic pre-treatment. (B) Secretion of granzyme B and perforin by unstimulated NK cells (Control) and A . fumigatus -stimulated NK cells depending (6 h) on the fungal strain and its enzymatic pre-treatment. (A-B) N = 6 independent donors. Repeated measures (RM) one-way analysis of variance (ANOVA) with Dunnett’s post-hoc test versus Control, i.e., unstimulated NK cells (asterisks). Additionally, enzymatic pre-treatments of A . fumigatus WT and the Δ agd3 mutant, respectively, were compared using RM one-way ANOVA with Dunnett’s post-hoc test versus no enzymatic pre-treatment (hash signs). Columns and error bars indicate means and standard deviations, respectively. */# p < 0.05, **/## p < 0.01, ***/### p < 0.001.

Article Snippet: Hyphae were incubated with soluble CD56 (5 μg/mL, R&D Systems, Minneapolis, MN, USA, Cat#2408-NC) or BSA (control, 5 μg/mL, Roth, Karlsruhe, Germany, #0163.2) in colorless RPMI for 2 h at 37°C, 5% CO 2 .

Techniques: Expressing, Control, Mutagenesis

Journal: Advanced biology

Article Title: CellMag-CARWash: A High Throughput Droplet Microfluidic Device for Live Cell Isolation and Single Cell Applications.

doi: 10.1002/adbi.202400066

Figure Lengend Snippet: Figure 2. Attachment of 2.8 μm Dynabeads to a desired cell population within a mixed cell population. A) Anti-CD56 coated Dynabeads attached to NK cells in a mixture of NK cells and T cells. i) Images of Dynabeads attached to NK cells (red). T cells are shown in blue. Scale bars indicate 20 μm. ii) Percentage of NK cells with various amounts of anti-CD56 Dynabeads. N = 3 iii) Percentage of T cells with various amounts of anti-CD56 coated Dynabeads. N = 3 B) Anti-EpCAM/ anti-EGFR/ anti-CD133 coated Dynabeads attached to MCF7 GFP cells in a mixture of MCF7 GFP cells and NK cells. i) Images of Dynabeads attached to MCF7 GFP cells (green). Scale bars indicate 20 μm. ii) Percentage of MCF7 GFP cell population with various amounts of anti-EpCAM/ anti-EGFR/ anti-CD133 coated Dynabeads. N = 4 iii) Percentage of NK cell population with various amounts of anti-EpCAM/ anti-EGFR/ anti-CD133 coated Dynabeads. N = 4.

Article Snippet: For NK cell attachment, biotinylated anti-CD56 antibodies (R&D Systems, cat# BAF2408) were then incubated with the beads at room temperature for 30 min on a rocker, allowing the biotinylated antibodies to bind with the streptavidin-coated Dynabeads.

Techniques: