Journal: Journal for Immunotherapy of Cancer

Article Title: IL-10 secretion by CD8 T cells orchestrates early NK cell recruitment and antitumor immunity after anti-VEGFR-2/PD-1 therapy

doi: 10.1136/jitc-2026-015243

Figure Lengend Snippet: NK cells produce GM-CSF, which promotes macrophage maturation. ( A ) Tumor growth of CT26-bearing mice receiving treatment with Ig, the bitherapy (αPD-1+ αVEGFR2), the α-asialo-GM1 antibody or both the bitherapy and the α-asialo-GM1 (n=10–12). ( B, C ) Flow cytometry of the percentage of macrophages MHCII high ( B ) 9 days, ( C ) 1, 2 and 3 days post-treatment. ( D ) TNFα, IFNγ and GM-CSF concentration (pg/mL/g) in the supernatant of CT26 tumor, 1-day, 2-day, 3-day and 9-day post-treatment. ( E ) Tumor growth of CT26-bearing mice receiving the indicated treatment (n=5–7). ( F ) Flow cytometry of the percentage of MHCII high macrophages at D9 post-treatment in CT26 tumor. ( G ) GM-CSF concentration in the supernatant of NK cells sorted from CT26-bearing mice treated or not and cultured for 24 hours. ( H, I ) GM-CSF concentration in the supernatant of intratumor NK cells from CT26 tumor 9 day post-treatment treated with the bitherapy and ( H ) an α-asialo-GM1 or ( I ) an αCD8β. ( J ) Flow cytometry of the percentage of MHCII high macrophages at D9 post-treatment in CT26 tumors. Data representative of at least three independent experiments. The statistical tests used were ( A, E ) two-way ANOVA with Tukey’s test, ( B, F, H, I–J ) one-way ANOVA with Tukey’s test, ( G ) unpaired t-test, ( C, D ) multiple unpaired t-tests. Data represent mean±SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.000. ANOVA, analysis of variance; GM-CSF, granulocyte-macrophage colony-stimulating factor; IFNγ, interferon-gamma; MHC, major histocompatibility complex; NK, natural killer; ns, not significant; PD1, programmed cell death protein-1; TNFα, tumor necrosis factor-alpha; VEGFR, vascular endothelial growth factor receptor.

Article Snippet: Peritoneal macrophages were incubated with increasing concentrations of mouse GM-CSF (Miltenyi Biotec, 130–094-043) in complete RPMI.

Techniques: Flow Cytometry, Concentration Assay, Cell Culture, Immunopeptidomics

Journal: Journal for Immunotherapy of Cancer

Article Title: IL-10 secretion by CD8 T cells orchestrates early NK cell recruitment and antitumor immunity after anti-VEGFR-2/PD-1 therapy

doi: 10.1136/jitc-2026-015243

Figure Lengend Snippet: GM-CSF produced in the tumor microenvironment promotes the recruitment of CD8 T cells via the CXCR3 receptor. ( A ) Flow cytometry at D9 post-treatment of the percentage of Ki-67 in CD8 T cells in CT26 tumor. ( B ) CXCL11 concentration in the supernatant of CT26 tumor at D9 post-treatment. ( C ) Relative concentration of CXCL11 measured by ELISA in the supernatant of CT26 tumor at day 1, 2, 3, 4, 5 and 9 days post-treatment (n≥10). ( D ) Tumor growth of CT26-bearing mice (n=7) receiving the indicated treatment. ( E ) Flow cytometry of the number of CD8 T cells in CT26 tumors. ( F ) Analysis of gene expression by reverse transcription quantitative real-time PCR (RT-qPCR) of Cxcl11 on macrophages sorted from CT26-bearing mice 5 days after the treatment, treated or not and cultured for 24 hours. ( G ) CXCL11 concentration in the supernatant of CT26 tumor at D9 post-treatment, determined by ELISA. ( H, I ) Flow cytometry of the numbers of CD8 T cells in CT26 tumor receiving the indicated treatment. ( J, K ) CXCL11 concentration in the supernatant of CT26 tumor at D9 post-treatment. Data representative of at least three independent experiments. The statistic test used were ( A, B, C, F ) unpaired t-test, ( D ) two-way ANOVA with Tukey’s test, ( E, G, H, I, J, K ) one-way ANOVA test with Tukey’s test. Data represent mean±SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance; GM-CSF, granulocyte-macrophage colony-stimulating factor; ns, not significant; PD1, programmed cell death protein-1.

Article Snippet: Peritoneal macrophages were incubated with increasing concentrations of mouse GM-CSF (Miltenyi Biotec, 130–094-043) in complete RPMI.

Techniques: Produced, Flow Cytometry, Concentration Assay, Enzyme-linked Immunosorbent Assay, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Cell Culture

Journal: Journal for Immunotherapy of Cancer

Article Title: IL-10 secretion by CD8 T cells orchestrates early NK cell recruitment and antitumor immunity after anti-VEGFR-2/PD-1 therapy

doi: 10.1136/jitc-2026-015243

Figure Lengend Snippet: Dual therapy enhances Treg recruitment; targeting Tregs further amplifies treatment efficacy. ( A-D ) Flow cytometry analysis ( A ) at D1 and ( B ) at D9 post-treatment of the numbers of Tregs in CT26 tumors, ( C ) of the percentage of intratumor CXCR3-expressing Tregs in CT26 tumor at D9, ( D ) of the number of Tregs in CT26 tumor at D9. ( E ) Heat map of normalized (z-score) mRNA gene expression from CT26 tumor at D9 post treatment (n=5) as analyzed by NanoString. ( F-H ) CT26-bearing mice receiving the indicated treatment. Tumor growth ( F ) and survival ( G ) curve are shown (n=5–7), ( H ) rechallenge of cured mice with naive mice as control (n=5–8 mice/group). ( I-K ) Flow cytometry analysis at D9 post-treatment of the numbers of Tregs in CT26 tumors. Data representative of at least three independent experiments. The statistical tests used were ( A, B, C ) unpaired t-test, ( D, E, I–K ) one-way ANOVA test with Tukey’s test, ( F, H ) two-way ANOVA with Tukey’s test, and ( G ) Kaplan-Meier survival analysis. Data represent ( E ) median and for others, mean±SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance; GM-CSF, granulocyte-macrophage colony-stimulating factor; mRNA, messenger RNA; ns, not significant; PD1, programmed cell death protein-1; Treg, regulatory T cell.

Article Snippet: Peritoneal macrophages were incubated with increasing concentrations of mouse GM-CSF (Miltenyi Biotec, 130–094-043) in complete RPMI.

Techniques: Flow Cytometry, Expressing, Gene Expression, Control

Journal: Frontiers in Medicine

Article Title: Long-Lasting Immunity Against SARS-CoV-2: Dream or Reality?

doi: 10.3389/fmed.2021.770381

Figure Lengend Snippet: Cellular immune profile and T cell phenotypes are stable over time, SARS-CoV-2 specific antibody levels decrease and then remain stable over time. Immune cell counts and T cell phenotypes were assessed by flow cytometric analysis and serological ELISA assays were performed to measure antibody levels. Results are shown for our large cohort of COVID-19 recovered donors (total n = 206 samples). (A) Symptom onset of every donor's sample, subdivided into three time periods with respect to time after symptom onset. (B) T cell phenotypes for CD3 + , CD4 + , and CD8 + cells in percentages. (C,D) Cellular immune profile expressed as blood cell counts per microliter. (E) Levels of anti-SARS-CoV-2 Spike IgG antibodies expressed as RU/ml. (F) Levels of anti-SARS-CoV-2 Nucleocapsid IgG expressed as IgG ratios. (B–F) Values are presented over the course of three time periods: 0–26 weeks ( n = 193), 27–52 weeks ( n = 82), and 53+ weeks ( n = 24) post symptom onset. (B–F) Kruskal-Wallis test followed by Dunn's multiple comparison. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns = non-significant.

Article Snippet: Overlapping peptide pools against the following peptides were used for stimulation at a final concentration of 1 μg of each peptide/ml peptide pool: SARS-CoV-2 membrane protein (M, #130-126-703), nucleocapsid protein (N, #130-126-699), and CMV phosphoprotein 65 (pp65, #130-093-435) (all Miltenyi Biotec, Bergisch Gladbach, Germany) as well as SARS-CoV-2 proteins S1 and S2 (PM-WCPV-S), huCoV epitopes (strains 229E, #PM-229E-S-1 and OC43, #PM-OC43-S-1; S1 and S2), and antigens derived from human RSV (nucleoprotein, NP, #PM-HRSVB-NCPN) and IAV (matrix protein 1, MP1, PM-INFA-MP1-H1N1) (all JPT, Berlin, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: Frontiers in Medicine

Article Title: Long-Lasting Immunity Against SARS-CoV-2: Dream or Reality?

doi: 10.3389/fmed.2021.770381

Figure Lengend Snippet: T cell frequencies against SARS-CoV-2, huCoV OC43 and 229E, RSV, IAV, and CMV epitopes are detectable and variously stable over time. Specific T cell frequencies were assessed by IFN-γ-EliSpot. Results are shown for our large cohort of COVID-19 recovered donors (total n = 206 samples) over the course of three time periods: 0–26 weeks ( n = 193), 27–52 weeks ( n = 82), and 53+ weeks ( n = 24) post symptom onset. Values are expressed as spots per 10,000 CD3 + cells. (A) T cell frequencies against the peptide pool derived from SARS-CoV-2 Membrane glycoprotein (M). (B) T cell frequencies against the peptide pool derived from SARS-CoV-2 Nucleocapsid protein (N). (C) T cell frequencies against the peptide pools derived from SARS-CoV-2, huCoV OC43, and huCoV 229E Spike S1 and S2 subunit proteins (D) Correlation of T cell frequencies against SARS-CoV-2 Spike S1 and S2 subunits. R 2 = 0.6114; P -value < 0.0001. (E) T cell frequencies against the peptide pools derived from RSV Nucleocapsid phosphoprotein (NP) and H1N1 IAV Matrix protein 1 (MP1). (F) T cell frequencies against the peptide pool derived from CMV phosphoprotein 65 (pp65). Results are shown for seropositive donors only (n=84) over the course of three time periods: 0-26 weeks ( n = 67), 27–52 weeks ( n = 28), and 53+ weeks ( n = 12) post symptom onset. (A–C,E,F) Kruskal-Wallis test followed by Dunn's multiple comparison. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns = non-significant.

Article Snippet: Overlapping peptide pools against the following peptides were used for stimulation at a final concentration of 1 μg of each peptide/ml peptide pool: SARS-CoV-2 membrane protein (M, #130-126-703), nucleocapsid protein (N, #130-126-699), and CMV phosphoprotein 65 (pp65, #130-093-435) (all Miltenyi Biotec, Bergisch Gladbach, Germany) as well as SARS-CoV-2 proteins S1 and S2 (PM-WCPV-S), huCoV epitopes (strains 229E, #PM-229E-S-1 and OC43, #PM-OC43-S-1; S1 and S2), and antigens derived from human RSV (nucleoprotein, NP, #PM-HRSVB-NCPN) and IAV (matrix protein 1, MP1, PM-INFA-MP1-H1N1) (all JPT, Berlin, Germany).

Techniques: Enzyme-linked Immunospot, Derivative Assay, Membrane, Comparison

Journal: Frontiers in Medicine

Article Title: Long-Lasting Immunity Against SARS-CoV-2: Dream or Reality?

doi: 10.3389/fmed.2021.770381

Figure Lengend Snippet: In our small individual cohort, T cell frequencies either are stable or decrease to a stable level over time, antibody level progressions vary from strong declines to stable levels over time. Specific T cell frequencies were assessed by IFN-γ-EliSpot, intracellular cytokines were detected by flow cytometric analysis and antibody levels were measured by serological ELISA assay. Results are shown for our small cohort of COVID-19 recovered donors ( n = 18). (A–D) Individual T cell frequencies against peptide pools derived from SARS-CoV-2 M, N, S1, or S2. Only responding donors to each respective peptide pool are shown. (E) Cumulative individual T cell frequencies against peptide pools derived from SARS-CoV-2 M, N, S1, and S2. All donors in this cohort are shown. (A–E) Values are expressed as spots per 10,000 CD3 + cells. (F) Intracellular cytokine staining and FACS analysis of PBMCs after 5 h of stimulation with the indicated peptides. Results are presented as frequencies of IFN-γ − /TNF-α + (blue), IFN-γ + /TNF-α + (green), and IFN-γ + /TNF-α − (yellow) among total T cells as well as T effector memory (TEM: CD45RA − CD62L + ) cells and T central memory (TCM: CD45RA − CD62L − ) cells within CD4 + and CD8 + T cell subsets [ n = 8; CMV pp65: seropositive only ( n = 6)]. Overall distribution of T cell phenotypes are displayed in the bar graph on the right side (mean + SD). (G) Individual levels of anti-SARS-CoV-2 Spike IgG antibodies expressed as RU/ml (RU = Relative Units). (H) Individual levels of anti-SARS-CoV-2 Nucleocapsid IgG antibodies expressed as IgG Ratios. (A–E,G,H) Results are divided into three time periods: 0–26 weeks ( n = 18), 27–52 weeks ( n = 7), and 53+ weeks ( n = 18) post symptom onset.

Article Snippet: Overlapping peptide pools against the following peptides were used for stimulation at a final concentration of 1 μg of each peptide/ml peptide pool: SARS-CoV-2 membrane protein (M, #130-126-703), nucleocapsid protein (N, #130-126-699), and CMV phosphoprotein 65 (pp65, #130-093-435) (all Miltenyi Biotec, Bergisch Gladbach, Germany) as well as SARS-CoV-2 proteins S1 and S2 (PM-WCPV-S), huCoV epitopes (strains 229E, #PM-229E-S-1 and OC43, #PM-OC43-S-1; S1 and S2), and antigens derived from human RSV (nucleoprotein, NP, #PM-HRSVB-NCPN) and IAV (matrix protein 1, MP1, PM-INFA-MP1-H1N1) (all JPT, Berlin, Germany).

Techniques: Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay, Derivative Assay, Staining

Journal: Immunity

Article Title: The pre-exposure SARS-CoV-2-specific T cell repertoire determines the quality of the immune response to vaccination

doi: 10.1016/j.immuni.2022.08.003

Figure Lengend Snippet:

Article Snippet: PepTivator Influenza A (H1N1) MP1 , Miltenyi Biotec , Cat#130-097-285.

Techniques: Functional Assay, Recombinant, Staining, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Software

Journal: Cell Reports

Article Title: Unravelling the sex-specific diversity and functions of adrenal gland macrophages

doi: 10.1016/j.celrep.2022.110949

Figure Lengend Snippet:

Article Snippet: Mouse M-CSF , Miltenyi Biotec , Cat# 130-094-129.

Techniques: Control, Recombinant, Staining, Enzyme-linked Immunosorbent Assay, Epinephrine ELISA, RNA Extraction, Software

Journal: Thorax

Article Title: A serum marker for the activity of pulmonary fibrosis in patients with systemic sclerosis

doi: 10.1136/thx.2005.la0181

Figure Lengend Snippet: Figure 1 (A) TNFa concentrations in bronchoalveolar lavage (BAL) fluid from healthy control subjects (n = 26), subjects with non-steroid treated mild asthma (n = 42) and those with steroid treated severe asthma (n = 20). For presentation purposes, one outlier (6660 fg/ml) in the severe asthma group is omitted from the graph. TNFa was measured by ELISA. (B) TNFa gene expression in endobronchial biopsy specimens from 14 patients with mild asthma and 14 with severe asthma. Total RNA was extracted and analysed by reverse transcription quantitative PCR. Results are normalised to the 18S rRNA levels and are presented as relative TNFa levels which were calculated using the DDCT method. (C) Immunoreactive TNFa positive cell counts in endobronchial biopsy specimens (cells/mm2) from 42 subjects with mild asthma and 14 subjects with severe asthma. (D) Immunohistochemical staining and localisation of TNFa in submucosal cells in a bronchial biopsy specimen from a subject with severe asthma. The data in A, B and C are presented as box and whisker plots showing the median, interquartile range, and 95% confidence limits with individual data points superimposed. Data were analysed using the Mann-Whitney U test and significant differences are indicated.

Article Snippet: Immunohistochemical staining for TNFa was conducted as previously described18 19 using a murine monoclonal antibody against human TNFa (1:500, R & D Systems).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Staining, Whisker Assay, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Truncating NFKB1 variants cause combined NLRP3 inflammasome activation and type I interferon signaling and predispose to necrotizing fasciitis

doi: 10.1016/j.xcrm.2024.101503

Figure Lengend Snippet:

Article Snippet: Human GM-CSF , Miltenyi , Cat#130-093-862.

Techniques: Control, Recombinant, Lysis, Protease Inhibitor, Western Blot, Cell Culture, Molecular Weight, Staining, Sequencing, Modification, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, Transfection, Quantitation Assay, Mutagenesis, Software, CRISPR, Variant Assay, Cytometry

Journal: Frontiers in Immunology

Article Title: Growth hormone-releasing hormone antagonist MIA-602 inhibits inflammation induced by SARS-CoV-2 spike protein and bacterial lipopolysaccharide synergism in macrophages and human peripheral blood mononuclear cells

doi: 10.3389/fimmu.2023.1231363

Figure Lengend Snippet: Inhibitory effect of MIA-602 on cytokine and chemokine expression in THP-1-derived macrophages. The cells were either untreated (c, control) or exposed to S protein (S, 500 ng/ml), MIA-602 (MIA, 1 µM) or LPS (50 ng/ml) alone for 24 h or pretreated with LPS for 3 h and then up to 24 h with S protein, in the presence or absence of MIA-602, at the same concentrations. (A–E) mRNA levels of TNF-α, IL-1β, IL-6, IL-8 and MCP-1 assessed by real time-PCR and normalized to 18S rRNA. (F–H) TNF-α, IL-1β and IL-6 secretion measured by ELISA in cell conditioned medium. Results are means ± SEM. * P < 0.05; *** P < 0.001 vs. control (c); # P < 0.05, ## P < 0.01, ### P < 0.001 vs. LPS; § P < 0.05, §§ P < 0.01, §§§ P < 0.001 vs. LPS+S protein by one-way ANOVA and Tukey’s post-hoc test ( n = 4).

Article Snippet: PepTivator SARS-CoV-2 Spike protein (S protein, code: 130-126-700) was purchased from Miltenyi Biotec (Bologna, Italy).

Techniques: Expressing, Derivative Assay, Control, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Growth hormone-releasing hormone antagonist MIA-602 inhibits inflammation induced by SARS-CoV-2 spike protein and bacterial lipopolysaccharide synergism in macrophages and human peripheral blood mononuclear cells

doi: 10.3389/fimmu.2023.1231363

Figure Lengend Snippet: Inhibitory effect of MIA-602 on LPS+S protein-induced activation of inflammatory pathways in THP-1-derived macrophages. Representative Western blots for phosphorylated (p)-p65 (A) , p-STAT3 (B) , p-ERK1/2 (C) and p-JNK (D) assessed by Western blot in THP-1 cells either untreated (c, control) or pretreated for 3 h with LPS (50 ng/ml), then up to 24 h with either S protein (500 ng/ml), MIA-602 (1 µM) or both (top panel). Blots were reprobed with non-phosphorylated antibodies for normalization (middle panel) or GAPDH, used as internal control (bottom panel). Graphs show the densitometric analysis of phosphorylated proteins normalized to total proteins and reported as percentage of control (means ± SEM). *** P < 0.001 vs. c; # P < 0.05, ## P < 0.01 vs. LPS; § P < 0.05 and §§ P < 0.01 vs. LPS+S protein by one-way ANOVA and Tukey’s post-hoc test ( n = 3).

Article Snippet: PepTivator SARS-CoV-2 Spike protein (S protein, code: 130-126-700) was purchased from Miltenyi Biotec (Bologna, Italy).

Techniques: Activation Assay, Derivative Assay, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: Growth hormone-releasing hormone antagonist MIA-602 inhibits inflammation induced by SARS-CoV-2 spike protein and bacterial lipopolysaccharide synergism in macrophages and human peripheral blood mononuclear cells

doi: 10.3389/fimmu.2023.1231363

Figure Lengend Snippet: Inhibitory activity of MIA-602 on release and expression of inflammatory cytokines and chemokines in human PBMCs. The cells were either untreated (c, control) or pretreated for 3 h with LPS (10 ng/ml), then up to 24 h with either S protein (500 ng/ml), MIA-602 (1 µM) or both. TNF-α (A) , IL-1β (B) and IL-6 (C) secretion measured by ELISA in cell conditioned medium. (D) MCP-1 mRNA expression assessed by real time-PCR. Results are means ± SEM. *** P < 0.001 vs. c; # P < 0.05 and ### P < 0.001 vs. LPS; § P < 0.05 and §§§ P < 0.001 vs. LPS+S protein by one-way ANOVA and Tukey’s post-hoc test ( n = 7 for A-C, n = 6 for D) .

Article Snippet: PepTivator SARS-CoV-2 Spike protein (S protein, code: 130-126-700) was purchased from Miltenyi Biotec (Bologna, Italy).

Techniques: Activity Assay, Expressing, Control, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Growth hormone-releasing hormone antagonist MIA-602 inhibits inflammation induced by SARS-CoV-2 spike protein and bacterial lipopolysaccharide synergism in macrophages and human peripheral blood mononuclear cells

doi: 10.3389/fimmu.2023.1231363

Figure Lengend Snippet: MIA-602-induced protection against oxidative stress in PBMCs. The cells were either untreated (c, control) or pretreated for 3 h with LPS (10 ng/ml), then up to 24 h with either S protein (500 ng/ml), MIA-602 (1 µM) or both. (A) Representative images of ROS production assessed by flow cytometry using the Muse ® Oxidative Stress Kit. M1: ROS negative cells, M2: ROS-positive cells. (B) Graph showing the percentage of ROS positive cells after the different treatments. Results are means ± SEM. *** P < 0.001 vs. control (c); ## P < 0.01 vs. LPS; § P < 0.05, vs. LPS+S protein by one-way ANOVA and Tukey’s post-hoc test ( n = 4). COX-2 (C) and iNOS (D) protein levels assessed by Western blot (top panels). Blots were reprobed with antibody for GAPDH for normalization (bottom panels). (E) Representative gelatin zymography of MMP9 activity in PBMC conditioned medium. Graph values show the densitometric analysis of MMP9, reported as percentage of control. Results are means ± SEM. *** P < 0.001 vs. control (c); ### P < 0.001 vs. LPS; §§§ P < 0.001 vs. LPS+S protein by one-way ANOVA and Tukey’s post-hoc test ( n = 6).

Article Snippet: PepTivator SARS-CoV-2 Spike protein (S protein, code: 130-126-700) was purchased from Miltenyi Biotec (Bologna, Italy).

Techniques: Control, Flow Cytometry, Western Blot, Zymography, Activity Assay

Journal: Frontiers in Immunology

Article Title: Growth hormone-releasing hormone antagonist MIA-602 inhibits inflammation induced by SARS-CoV-2 spike protein and bacterial lipopolysaccharide synergism in macrophages and human peripheral blood mononuclear cells

doi: 10.3389/fimmu.2023.1231363

Figure Lengend Snippet: Inhibitory effect of MIA-602 on LPS+S protein-induced translocation and activation of NF-кB in PBMCs. The cells were either untreated (c, control) or pretreated for 3 h with LPS (10 ng/ml), then up to 24 h with S protein (500 ng/ml) or S protein+MIA-602 (1 µM). (A) Representative fluorescence microscopy images showing cells stained for NF-кB p65 (green). Nuclei were counterstained with DAPI (blue). Insets show higher magnification (scale bar: 200 μm). (B) Representative Western blot for phosphorylated (p)-p65. Blots were reprobed with non-phosphorylated p65 for normalization. Graph shows the densitometric analysis, reported as percentage of control (means ± SEM). *** P < 0.001 vs. c; # P < 0.05, ### P < 0.001 vs. LPS, and §§§ P < 0.01 vs. LPS+S protein by one-way ANOVA and Tukey’s post-hoc test ( n = 3).

Article Snippet: PepTivator SARS-CoV-2 Spike protein (S protein, code: 130-126-700) was purchased from Miltenyi Biotec (Bologna, Italy).

Techniques: Translocation Assay, Activation Assay, Control, Fluorescence, Microscopy, Staining, Western Blot

Journal: Cell Reports Medicine

Article Title: IL-21/IL-21R signaling renders acute myeloid leukemia stem cells more susceptible to cytarabine treatment and CAR T cell therapy

doi: 10.1016/j.xcrm.2024.101826

Figure Lengend Snippet:

Article Snippet: Human recombinant IL-15 , Miltenyi , Cat# 130-095-760.

Techniques: Control, Recombinant, Virus, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Plasmid Preparation, Software, Phospho-proteomics