Journal: Cells

Article Title: Cancer Immunomodulatory Effect of Bidens pilosa L. in Mice: Suppression of Tumor-Associated Macrophages and Regulatory T Cells

doi: 10.3390/cells15020126

Figure Lengend Snippet: Expression levels of mRNAs and proteins associated with TAMs and Tregs in BPA -treated mouse tumors. ( A ) Quantification of mRNA by qRT-PCR for TAM/Treg-associated tumor-promoting factors (Arg-1, YM-1, CCL2, CCL22, Foxp3, CD25, TGF-β, CXCR3, CCR4, CCR8, CCR10) and tumor-inhibitory factors (TNF-α, IL-1β, iNOS, CXCL10, CCR7). Data were normalized to 18 S rRNA and are shown relative to the model group (baseline control). ( B ) Representative immunoblots of tumor tissue for TAM- and Treg-associated proteins. Pro-tumor: CD206, Foxp3, CD25, PD-1, PD-L1; Anti-tumor: CD80, CD4, GITR. ( C ) Quantification of protein expression from Western blots. Values were normalized to α-tubulin, β-actin, or GAPDH as indicated. Statistical significance is denoted as follows ( n = 6): compared to the model group, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: To block Fc receptors, cells were incubated with anti-CD16/32 (Elabscience, Cat. E-AB-F0997A, Wuhan, China) for 10 min. After washing, cells were stained with PerCP/Cyanine 5.5 anti-mouse F4/80 (Elabscience, Cat. E-AB-F0995J), FITC anti-mouse CD206 antibody (Elabscience, Cat. E-AB-F1135C), and APC anti-mouse CD80 antibody (Elabscience, Cat. E-AB-F0992E) at 4 °C for 30 min.

Techniques: Expressing, Quantitative RT-PCR, Control, Western Blot

Journal: Theranostics

Article Title: Macrophage membrane-biomimetic cinnamaldehyde nanomedicine ameliorates inflammatory bowel disease by suppressing macrophage M1 polarization

doi: 10.7150/thno.124748

Figure Lengend Snippet: CMA skews macrophage polarization toward M2 while attenuating M1 differentiation. (A) Chemical structural formula of CMA. (B) RAW 264.7 cells were treated with CMA (0-80 μM) for 24 h, followed by incubated with MTT (0.5 mg/mL) for 4 h (n = 5). (C) Schematic illustration of macrophage polarization mediated by different inducers. (D-F) The mRNA levels of M1 markers ( Nos2, Cd80, and Tnf ) in RAW 264.7 macrophages pretreated with CMA (20 μM) for 1 h, followed by co-treatment with LPS (1 µg/mL) for 24 h (n = 3). (G, H) The mRNA levels of M2 markers ( Arg1 and Cd206 ) in RAW 264.7 macrophages pretreated CMA (20 μM) for 1 h, followed by co-treatment with IL-4 (20 ng/mL) for 24 h (n = 3). (I, J) Flow cytometry analysis and quantification of CD80 protein levels in M1 macrophages treated with or without CMA (20 μM) for 24 h (n = 3). (K, L) Flow cytometry analysis and quantification of CD206 in M2 macrophages treated with or without CMA (20 μM) for 24 h (n = 3). (M-O) Effects of CMA treatment on NO levels (n = 4) and cell migration (n = 3) in RAW 264.7 macrophages. Scale bar: 100 μm. Data presented as mean ± SD, *P < 0.05, **P < 0.01.

Article Snippet: Then, the Fc receptors were blocked by treating with CD16/32 antibody (Elabscience Biotechnology Co., Ltd., Cat. E-AB-F0997A) for 10 min. For polarization analysis, M1 macrophages were stained with fluorescein isothiocyanate (FITC)-conjugated CD80 antibody (Proteintech, Cat. FITC-65076), while M2 macrophages were labeled with FITC-conjugated CD206 antibody (Elabscience, Cat. E-AB-F1135C).

Techniques: Incubation, Flow Cytometry, Migration

Journal: Theranostics

Article Title: Macrophage membrane-biomimetic cinnamaldehyde nanomedicine ameliorates inflammatory bowel disease by suppressing macrophage M1 polarization

doi: 10.7150/thno.124748

Figure Lengend Snippet: MM@CMANP modulates macrophages polarization and enhances autophagy in IBD mice. (A-D) Protein concentrations of TNF-α, IL-6, IL-1β and IL-10 in mouse serum were detected by ELISA assay (n = 5). (E) Representative immunohistochemical staining images of M1 marker CD80 and M2marker CD206 in mouse colon tissues. Scale bar: 50 μm. (F, G) Representative flow cytometry plots showing the frequencies of CD80 + (f) and CD206 + (g) cells gated on colonic lamina propria macrophages (F4/80 + and CD11b + ) from Control, DSS, DSS + CMANP, and DSS + MM@CMANP mice. (H, I) Quantification of the percentages of CD80 + (h) and CD206 + (i) cells within F4/80 + CD11b + macrophages. (J) Relative M1/M2 ratio (or composition) calculated based on CD80 + and CD206 + macrophage populations (n = 3). (K, L) Representative immunofluorescence images and relative quantitative statistics of autophagosome marker LC3B in mouse colon tissues (n = 5). Scale bar: 50 μm. Data presented as mean ± SD, *P < 0.05, **P < 0.01.

Article Snippet: Then, the Fc receptors were blocked by treating with CD16/32 antibody (Elabscience Biotechnology Co., Ltd., Cat. E-AB-F0997A) for 10 min. For polarization analysis, M1 macrophages were stained with fluorescein isothiocyanate (FITC)-conjugated CD80 antibody (Proteintech, Cat. FITC-65076), while M2 macrophages were labeled with FITC-conjugated CD206 antibody (Elabscience, Cat. E-AB-F1135C).

Techniques: Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Marker, Flow Cytometry, Control, Immunofluorescence