Journal: Frontiers in Immunology

Article Title: Nocardia rubra cell wall skeleton-induced MARCO expression: implications for improved phagocytosis and cytokine secretion in tumor-associated macrophages

doi: 10.3389/fimmu.2026.1611476

Figure Lengend Snippet: Nr-CWS affects MARCO expression through TLR4. The expression of TLR4 significantly increased after Nr-CWS treatment [ (D-G) , scale bars = 100 μm], while the expression TLR2 was unchanged (A-C) . Subsequent inhibition of TLR4 expression resulted in decreased levels of MARCO [ (H-J) , scale bars = 100 μm]. n = 3, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The following commercial antibodies (vendor, catalog number and dilution) utilized for western blot and immunohistochemical staining were employed in accordance with the manufacturers’ guidelines: mouse anti-CD68 antibody (Abcam, ab201340, 1:200), rabbit anti-MARCO antibody (Abcam, ab231046, 1:1000 for western blot and 1:100 for immunohistochemistry), rabbit anti-CD163 antibody (Abways Technology, CY6845, 1:500), mouse anti-CD86 antibody (Proteintech, 68674-2-Ig, 1:5000), mouse anti-TLR4 antibody (Santa Cruz Biotechnology, sc-293072, 1:1000 for western blot and 1:200 for immunohistochemistry), rabbit anti-TLR2 antibody(Abways Technology, CY5102, 1:2000), mouse anti-GAPDH antibody (Proteintech, 6004-1-Ig, 1:50000), HRP-goat anti-mouse recombinant secondary antibody (H+L)(Proteintech, RGAM001, 1:5000), goat anti-mouse IgG (H+L) Alexa Fluor 594 (Abways Technology, AB0152, 1:300), HRP-goat anti-rabbit recombinant secondary antibody (H+L) (Proteintech, RGAR001, 1:5000), goat anti-rabbit IgG (H+L) secondary antibody DyLightTM 594 (Report Biotech, S7002, 1:500), goat anti-mouse IgG (H+L) secondary antibody DyLightTM 488 (Report Biotech, S6001, 1:500), goat anti-mouse IgG (H+L) secondary antibody DyLightTM 594 (Abways Technology, AB0152, 1:500).

Techniques: Expressing, Inhibition

Journal: ACS Nano

Article Title: Microplastic-Induced Macrophage Dysfunction Drives Lung Tumor Progression through Glutathione Imbalance

doi: 10.1021/acsnano.5c15425

Figure Lengend Snippet: MPs induce inflammation by recognizing TLR2/4 in BMDMs. (A) Representative Western blot images of BMDMs treated with MPs or LPS (positive control) for 24 h. TLR2, TLR4, TIRAP, MyD88, TRAF6, P-JNK, and P-c-Jun protein expression levels, analyzed by Western blotting. β-actin expression was used as a loading CTL. (B) Relative mRNA expression levels of Il-1β , Il-6 , and Tnf-α , measured in the presence of MP. Gapdh was used for normalization. (C) Representative flow cytometry plots presenting BMDM M1 and M2 polarization upon MP treatment. The M1 and M2 macrophage populations were quantified as CD86 + CD206 – and CD86 – CD206 + , respectively. (D) Representative immunofluorescence images of BMDMs treated with or without MPs for 24 h, stained with MPO (green), CitH3 (red), and DAPI (blue). Scale bars, 10 μm. Fluorescence intensity-based MPO and CitH3 quantification in BMDMs. a.u., arbitrary unit. The data represent at least three independent experiments and are expressed as the mean ± SEM, using one-way ANOVA. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: Each membrane was incubated overnight at 4 °C with primary antibodies against TLR2 (Cell Signaling Technology, 13744S), TLR4 (Proteintech, 66350–1-Ig), TIRAP (Cell Signaling Technology, 13077), MyD88 (Cell Signaling Technology, 4283), TRAF6 (Cell Signaling Technology, 67591), P-JNK (Cell Signaling Technology, 9251), P-c-Jun (Cell Signaling Technology, 3270), NRF2 (Cell Signaling Technology, 12721), GCLM (Abcam, ab126704), SLC7A11 (Novus Biologicals, NB300–318), NQO1 (Santa Cruz Biotechnology, sc-32793), GCLC (Cell Signaling Technology, 52183), GPX1 (R&D systems, AF3798), GPX3 (R&D systems, AF4199), GPX4 (R&D systems, MAB5457), FTL (Santa Cruz Biotechnology, sc-390558;), FTH1 (Santa Cruz Biotechnology, sc-376594), and β-actin (Cell Signaling Technology, 8457).

Techniques: Western Blot, Positive Control, Expressing, Flow Cytometry, Immunofluorescence, Staining, Fluorescence

Journal: Cancers

Article Title: Crosstalk of Tumor-Derived Extracellular Vesicles with Immune Recipient Cells and Cancer Metastasis

doi: 10.3390/cancers18020196

Figure Lengend Snippet: Activation of the NF-κb pathway by MTEX in THP-1 recipient cells. In ( A ), NF-κb phosphorylation, p38 phosphorylation and a decrease in expression levels of TLR2 in THP-1 cells co-incubated with MTEX. In ( B ), Expression levels of p-NF-κb and p-p38 are decreased in the presence of anti-TLR2 mAb ( right ).

Article Snippet: In selected experiments, anti-TLR2 Ab (0.5 μg/mL; Cell Signaling, Boston, MA, USA 12276) or Bafilomycin A1 (BafA1, 10 nM, Sigma, 131793) were added to wells containing THP-1 cells and EVs.

Techniques: Activation Assay, Phospho-proteomics, Expressing, Incubation