Journal: Frontiers in Immunology

Article Title: Decidual stromal cells drive CD16 + macrophages towards an immunoregulatory phenotype via extracellular matrix-adhesion molecule interaction during early pregnancy

doi: 10.3389/fimmu.2025.1747323

Figure Lengend Snippet: Exogenous COL4A1, OPN and HA promote development of immunomodulatory CD16 + Mφ. (A, B) After 48 hours of stimulation with exogenous COL4A1 (10 ug/mL), OPN (1 ug/mL) or vehicles, the expression levels of CD16 on U937-induced Mφ ( A ; n=5) and the polarization-related molecules CD86, CD209 and CD206 on CD16 - or CD16 + Mφ ( B ; n=5) were detected by flow cytometry. (C, D) After 48 hours of stimulation with exogenous HA (0, 50, 100 μM), the expression levels of CD16 on U937-induced Mφ ( C ; n=4) and CD86, CD209 and CD206 on CD16 - or CD16 + Mφ ( D ; n=4) were detected by flow cytometry. The data are presented as the mean ± SEM; oneway ANOVA test or two-tailed Student’s t-test; *P < 0.05, **P < 0.01, ****P< 0.0001, NS, no significant difference.

Article Snippet: Human monocyte cell line U937 from American Type Culture Collection (ATCC, CRL-3253) was cultured with RPMI-1640 medium (HyClone, SH30027.01) containing 10% FBS.

Techniques: Expressing, Flow Cytometry, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Decidual stromal cells drive CD16 + macrophages towards an immunoregulatory phenotype via extracellular matrix-adhesion molecule interaction during early pregnancy

doi: 10.3389/fimmu.2025.1747323

Figure Lengend Snippet: CD16 + Mφ in the co-culture system are suppressed with the inhibition of COL4A1, OPN and HA in DSCs. (A) DSCs were transfected by plasmid of siRNA targeting COL4A1 (si COL4A1 ; n=3), SPP1 (si SPP1 ; n=3) or control plasmids (n=3) for 72 hours and the efficacy was verified by RT-qPCR. (B-D) After co-cultured with COL4A1 -silenced DSCs, SPP1 -silenced DSCs or control DSCs for 48 hours, CD16 expression of U937-induced Mφ ( B, C ; n=4) and the polarization markers (CD86, CD209 and CD206) of CD16 - or CD16 + macrophages ( D ; n=4) were explored by flow cytometry. (E, F) After co-cultured with 4-MU (a hyaluronic Acid synthesis inhibitor, 500 μM) or vehicle treated DSCs for 48 hours, CD16 expression of U937-induced Mφ ( E ; n=6) and the polarization markers of CD16 - or CD16 + macrophages ( F ; n=6) were explored by flow cytometry. The data are presented as the mean ± SEM; oneway ANOVA test or two-tailed Student’s t -test; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns, NS significant difference.

Article Snippet: Human monocyte cell line U937 from American Type Culture Collection (ATCC, CRL-3253) was cultured with RPMI-1640 medium (HyClone, SH30027.01) containing 10% FBS.

Techniques: Co-Culture Assay, Inhibition, Transfection, Plasmid Preparation, Control, Quantitative RT-PCR, Cell Culture, Expressing, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: A Pseudomonas aeruginosa quorum-sensing metabolite manipulates macrophage ferroptosis through a methylation pathway

doi: 10.1038/s41467-025-65142-y

Figure Lengend Snippet: A Growth curves of PAO1, Δ pqsR , and Δ pqsR + strains. B Lactate dehydrogenase (LDH) assay for cell death in RAW264.7 macrophages infected with PAO1, Δ pqsR , and Δ pqsR + (Δ pqsR mutant expressing pqsR from a rescue construct pMiniCTX1- pqsR ) Bac (bacteria cultures) or Sup (cell-free supernatants). Multiplicity of infection (MOI) = 100. Data represent the means ± SD ( n = 3, two-way ANOVA with Tukey’s multiple comparisons test). C ELISA quantification of PQS concentrations measured in PAO1, Δ pqsR , and Δ pqsR + supernatants. Data represent the means ± SD ( n = 3, one-way ANOVA with Tukey’s multiple comparisons test). D LDH assay for cell death in RAW264.7 macrophages treated with different concentrations of PQS for 24 h, untreated macrophages (Ctrl), and macrophages treated with DMSO as the negative control. Data represent the means ± SD ( n = 3, one-way ANOVA with Tukey’s multiple comparisons test). E CCK-8 assay for cell viability in RAW264.7 macrophages treated with different concentrations of PQS for 24 h; IC 50 (PQS) = 14.98 μg/mL. Data represent the means ± SD ( n = 3). F Microscopic observation of the effect of concentration of IC 50 (PQS) exposure for 24 h on RAW264.7 macrophage morphology. Scale bar = 100 µm. G Transcription electron microscopy (TEM) observation of the effect of 10 μg/mL PQS exposure on RAW264.7 macrophage morphology. Nu: cell nucleus, Au-L: autolysosome. 15/40KK: × 15000/ 40,000 magnifications. Scale bar = 2/0.5 μm. H CCK-8 assay for cell viability in BMDM, THP-1, U937, A549, and BEAS-2B cells treated with 10 μg/mL PQS for 24 h. Data represent the means ± SD ( n = 3). Source data are provided as a Source Data file.

Article Snippet: The RAW264.7 murine macrophages (ATCC ® TIB-71 TM ), THP-1 human monocyte (ATCC ® TIB-202 TM ), U937 human leukaemia (ATCC ® CRL-3253 TM ), A549 human epithelium (ATCC®CCL-185TM), and BEAS-2B human bronchial epithelium (ATCC ® CRL-3588 TM ) cell lines were grown in Dulbecco’s Modified Eagle (DMEM) or Roswell Park Memorial Institute (RPMI)-1640 medium supplemented with 10% foetal bovine serum (FBS) and 1% penicillin/streptomycin (PS) (Gibco, Texas, USA), followed by incubation at 37 °C in a 5% CO 2 incubator.

Techniques: Lactate Dehydrogenase Assay, Infection, Mutagenesis, Expressing, Construct, Bacteria, Enzyme-linked Immunosorbent Assay, Negative Control, CCK-8 Assay, Concentration Assay, Electron Microscopy