Journal: bioRxiv

Article Title: Antimitotic chemotherapy promotes tumor NF-kB secretory phenotype and immunosuppressive CXCR2+ neutrophils chemotaxis in triple-negative breast cancers

doi: 10.1101/2024.12.21.629876

Figure Lengend Snippet: A: PCA analysis of 4T1-Luc2 CRISPR CTL and CRISPR STING treated or not by paclitaxel and analyzed by 3’Seq RNA Profiling. B, C: Volcano plots representing differentially up- and downregulated expressed under paclitaxel treatment in 4T1-Luc2 CRISPR CTL and CRISPR STING , respectively. D, E: Signaling pathway signatures up- or downregulated by paclitaxel in 4T1-Luc2 CRISPR CTL and CRISPR STING , respectively. F, G: Concentration in CCL5 and CXCL1 by ELISA in the culture medium of 4T1-Luc2 CRISPR CTL and CRISPR STING treated or not by paclitaxel. H: Concentration in CXCL1 by ELISA spot in the culture medium of paclitaxel-treated 4T1-Luc2 CRISPR CTL and CRISPR STING under treatment with AS602868 or not.

Article Snippet: For ELISA assays, levels of different chemokines in cell supernatants were determined according to the manufacturer’s protocols using following kits: CXCL1 (Mouse CXCL1 Sandwich ELISA Kit, Proteintech) and CCL5 (Mouse CCL5 Sandwich ELISA Kit, Proteintech).

Techniques: CRISPR, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Antimitotic chemotherapy promotes tumor NF-kB secretory phenotype and immunosuppressive CXCR2+ neutrophils chemotaxis in triple-negative breast cancers

doi: 10.1101/2024.12.21.629876

Figure Lengend Snippet: A: Impact of STING on paclitaxel-induced transcriptional expression (3’SRP) of Cxcl1, Cxcl5, Ccl5 and Ccl20 comparing fold-change expression (paclitaxel versus untreated) in both CRISPR CTL and CRISPR STING . B: Transcriptional expression fold change of Cxcl1, Cxcl5, Ccl5 and Ccl20 in CRISPR CTL and CRISPR STING under paclitaxel or not. C : Expression of CXCL1 and CCL5 by human TNBC cells using scRNAseq analysis from Broad Institute ( https://singlecell.broadinstitute.org/single_cell/study/SCP1106/stromal-cell-diversity-associated-with-immune-evasion-in-human-triple-negative-breast-cancer ).

Article Snippet: For ELISA assays, levels of different chemokines in cell supernatants were determined according to the manufacturer’s protocols using following kits: CXCL1 (Mouse CXCL1 Sandwich ELISA Kit, Proteintech) and CCL5 (Mouse CCL5 Sandwich ELISA Kit, Proteintech).

Techniques: Expressing, CRISPR

Journal: Regenerative biomaterials

Article Title: Concentrated growth factor regulates the macrophage-mediated immune response.

doi: 10.1093/rb/rbab049

Figure Lengend Snippet: Figure 4. CGF extract modulated macrophage cytokine secretion. (A) The supernatants of CCM-treated macrophages were screened using a human cytokine anti- body array. (B) Representative cytokines are presented. (C) A GO analysis of differentially expressed cytokines enriched in BP terms after 20% CCM treatment. (D) Enriched MF categories. (E) The mRNA expression of differentially expressed cytokines (IL-1b, IL-7, RANTES and MCP-1) in CCM-treated macrophages was ana- lyzed using q-PCR. (F) The secretion of IL-1b and RANTES by CCM-treated macrophages was further confirmed using ELISAs. The quantitative data are presented as the mean 6 SD (n ¼ 3) and were analyzed using one-way analysis of variance (ANOVA) and post hoc Dunnett tests: *P< 0.05, **P< 0.01 and ***P < 0.001.

Article Snippet: The supernatants were carefully subjected to Enzyme-linked immunosorbent assays (ELISAs) for RANTES (KE00093, Proteintech Group, China) and IL-1b (KE00021, Proteintech Group, China) using a standard protocol.

Techniques: Expressing

Journal: Regenerative biomaterials

Article Title: Concentrated growth factor regulates the macrophage-mediated immune response.

doi: 10.1093/rb/rbab049

Figure Lengend Snippet: Figure 5. CGF extract regulated the macrophage-mediated immune response via the AKT pathway. (A) A western blot analysis of the proteins involved in the PI3K/AKT (p-AKT, AKT and PI3K) and JAK/STAT (p-JAK, JAK, p-STAT3 and STAT3) signaling pathways in CCM-treated THP-1 macrophages. AKT phosphorylation was increased, but no significant differences were observed in the JAK/STAT pathway (n ¼ 3). (B) The effect of an AKT inhibitor on AKT pathway inhibition was analyzed using western blotting (n ¼ 3). (C) Representative images of adherent THP-1 macrophages cultured in CCM with or without AKT inhibitor were analyzed using ImageJ software (n ¼ 5). (D) The secretion of IL-1b and RANTES by THP-1 macrophages were assayed using ELISAs (n ¼ 3). (E) Expression of the M2 sub- type marker CD163 was detected using immunofluorescence staining (n ¼ 3). The data are presented as the mean 6 SD and were analyzed using one-way analy- sis of variance (ANOVA) and post hoc tests: *P < 0.05, **P < 0.01 and ***P < 0.001.

Article Snippet: The supernatants were carefully subjected to Enzyme-linked immunosorbent assays (ELISAs) for RANTES (KE00093, Proteintech Group, China) and IL-1b (KE00021, Proteintech Group, China) using a standard protocol.

Techniques: Western Blot, Protein-Protein interactions, Phospho-proteomics, Inhibition, Cell Culture, Software, Expressing, Marker, Staining

Journal: Nature communications

Article Title: Asthma reduces glioma formation by T cell decorin-mediated inhibition of microglia.

doi: 10.1038/s41467-021-27455-6

Figure Lengend Snippet: Fig. 3 Asthma reduces T cell induction of Ccl5 in microglia. a Schematic representation of the mouse Nf1OPG neuron-immune-cancer cell axis in the setting of asthma. b No difference in optic nerve Mdk RNA expression was observed between OVA- (n = 3) or HDM- (n = 3) treated mice and controls (n = 6). Bar graphs represent the means ± SEM of independent biological samples. One-way ANOVA with Bonferroni post hoc correction. c Ccl4 levels are similarly induced with increasing midkine concentrations (0–100 ng/ml) in CD3+ T cells from OVA- and PBS-treated mice. Data are presented as the means ± SEM of n = 3 independent biological samples. One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. d, e Similarly increased CD8+, relative to CD4+, T cell content was detected in the optic nerves of Nf1OPG mice treated with PBS (n = 8), OVA (n = 8) or HDM (n = 8). One-way ANOVA with Bonferroni post hoc correction. Activated T cell medium from (f) OVA- (n = 4) and g HDM- (n = 4) treated mice induced lower levels of microglial Ccl5 relative to PBS-treated Nf1OPG mice. Two-tailed Student’s t test. Similar reductions of Ccl5 expression were observed in CD4+ and CD8+ T cells from (h) OVA- (n = 3) and (i) HDM- (n = 3) treated mice relative to PBS-treated controls (n = 3). One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. d Scale bars, 40 µm. From left to right in each panel: b ns; c ns; ns; ns; ns; e P < 0.0001, P = 0.0022, P = 0.0011; f P = 0.0008; g P = 0.0015; h P = 0.0186, P = 0.0054; i P = 0.0224, P = 0.0007.

Article Snippet: In total, 5 × 105 microglia were grown in T cell-conditioned media (TCM) for 48 h8,9, followed Ccl5 (R&D Systems, MMR00) and decorin (Abcam, ab155454) determinations by ELISA.

Techniques: RNA Expression, Two Tailed Test, Expressing