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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.

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

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.

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