neutralization antibody experiment  (Proteintech)

Journal: Nature communications

Article Title: Type I interferon drives T cell cytotoxicity by upregulation of interferon regulatory factor 7 in autoimmune kidney diseases in mice.

doi: 10.1038/s41467-025-59819-7

Figure Lengend Snippet: Fig. 5 | IFN-I signaling in T cells drives GzmB production and renal tissue damage. a CD8+ and CD4+ T cells were isolated from the spleen of Ifnar1 KO or wild- type mice, and transferred into Rag1 KO mice (WT, n = 8; KO, n = 9). b Representative photographs of PAS staining of the kidney. Quantification is also shown (WT, n = 8; KO, n = 9). Representative histograms and bar graphs showing the MFI of IRF7 in CD8+ T cells (c) and CD4+ T cells (d) isolated from the kidney (WT, n = 8; KO, n = 9). e Representative flow cytometry plots and a bar graph illustrating the production of GzmB in CD8+ T cells (WT, n = 8; KO, n = 9). f Bar graph showing the production of GzmB in CD4+ T cells (WT, n = 8; KO, n = 9). g cGN was induced in mice, and mice were injected with anti-IFNAR1 Ab (n = 10) or isotype control (n = 9)

Article Snippet: For the IFNAR1 neutralization experiment, we injected 100μg of InVivoMAb anti-mouse IFNAR-1 antibody (BioXCell, BE0241) or InVivoMAb anti-mouse IgG1 isotype control antibody (BioXCell, BE0083) on days 0, 3, 6, and 9 of the cGN.

Techniques: Isolation, Staining, Cytometry, Injection, Control

Journal: Oncoimmunology

Article Title: Cancer-cell derived S100A11 promotes macrophage recruitment in ER+ breast cancer

doi: 10.1080/2162402X.2024.2429186

Figure Lengend Snippet: ER+ breast cancer cell-derived S100A11 promotes monocyte recruitment in a 3D collagen matrix. (a) ELISA assay to quantify S100A11 secretion level by ER+ breast cancer cell lines. T47D cells exhibit higher S100A11 levels compared to MM330 or BT483 ( n = 2 biological replicates). (b) 3D collagen matrix experiment to quantify CD14+ monocyte recruitment by cancer cells. T47D cancer cells recruited the highest number of monocytes compared to MM330 or BT483 (biological replicates: n = 4 for T47D, BT483; n = 3 for MM330). (c) Time-lapse images of monocyte infiltration into 3D collagen matrices embedded with T47D for 24 h. Red dotted lines indicate the interface between media and collagen matrices. Orange arrows indicate the infiltrated CD14+ monocytes. Neutralization of S100A11 using a blocking antibody decreased the number of recruited monocytes. Scale bars, 100 µm. (d) Quantification of recruited CD14+ monocytes in images shown in (c) ( n = 3 biological replicates). (e) Normalized number of CD14+ monocytes that infiltrated into the collagen matrices embedded with control siRNA or S100A11 siRNA-treated T47D cells ( n = 4 biological replicates). (f) Visualization of cancer-monocyte interactions in organoids embedded in the collagen gel after 24 h under IgG control or anti-S100A11 conditions. Scale bars, 50 µm. Red: actin staining; Green: CMFDA monocyte staining; blue: nuclear staining. (G) Treatment with the S100A11 blocking antibody decreased monocyte recruitment toward breast cancer organoids in collagen matrices ( n = 3 biological replicates).

Article Snippet: For the neutralization antibody experiment, a S100A11 blocking antibody (Proteintech, Cat# 10237–1-AP) was added at a concentration of 1 μg/ml.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Neutralization, Blocking Assay, Control, Staining

Journal: Neuron

Article Title: PDGFRβ Cells Rapidly Relay Inflammatory Signal from the Circulatory System to Neurons via Chemokine CCL2.

doi: 10.1016/j.neuron.2018.08.030

Figure Lengend Snippet: Figure 7. LPS- or Poly(I:C)-Treated HBVP Extracellular Solution Increases Excitatory Synaptic Transmission and Neuronal Excitability in a CCL2-Dependent Manner (A) Concentration of CCL2 in HBVP extracellular solution (ECS) after LPS or Poly(I:C) treatment (13–14 independent samples per condition; Kruskal-Wallis test, Dunn’s post hoc test). (B and C) The effect of Ctrl ECS, LPS-treated ECS, or Poly(I:C)-treated ECS on mEPSC frequency and amplitude of CA1 pyramidal neurons, representative traces (B) and summary data (C) (13–14 cells from 3 mice per group; Kruskal-Wallis test, Dunn’s post hoc test). (D and E) The effect of Ctrl ECS, LPS ECS, or Poly(I:C) ECS on firing responses to stepwise current injections in CA1 pyramidal neurons, representative traces (D) and summary data (E) (14–16 cells from 3 mice; two-way ANOVA, Bonferroni post hoc test). (F) Concentration of CCL2 in LPS-stimulated ECS after neutralization with human CCL2 antibody (4 independent samples per group; Kruskal-Wallis test, Dunn’s post hoc test). (G and H) The effect of Ctrl ECS, LPS ECS, or human CCL2 antibody neutralized LPS ECS (LPS ECS+ a-CCL2) on mEPSC frequency and amplitude, repre- sentative traces (G) and summary data (H) (14–15 cells from 2–4 mice; Kruskal-Wallis test, Dunn’s post hoc test). (I and J) The effect of Ctrl ECS, LPS ECS, or LPS ECS + a-CCL2 on firing responses to stepwise current injections, representative traces (I) and summary data (J) (17–22 cells from 2–4 mice; two-way ANOVA, Bonferroni post hoc test). (K and L) The effect of saline-treated ECS (ECS) or human CCL2 antibody neutralized saline-treated ECS (ECS + a-CCL2) on mEPSC frequency and amplitude, representative traces (K) and summary data (L) (13–14 cells from 3 mice; Mann-Whitney test). See also Figures S8 and S9.

Article Snippet: For the antibody neutralization experiment, human CCL2 antibody (R&D Systems, Cat# MAB679-500, 1:100) were added to the ECS and incubated at 37 C for 2 h before experiments.

Techniques: Transmission Assay, Concentration Assay, Neutralization, Saline, MANN-WHITNEY