Journal: Frontiers in Immunology

Article Title: Bufei Jiedu Formula enhances CD40 activation and macrophage polarization to eliminate intracellular MRSA persisters

doi: 10.3389/fimmu.2025.1623182

Figure Lengend Snippet: BFJD promotes the clearance of persistent MRSA infection and macrophage M1 polarization in vivo . (A) Schematic illustration of a 42-day long-term persistent MRSA infection model. Mice were inoculated with 1×10 8 CFU of MRSA in 0.2 ml of PBS via a lateral tail vein. At day 28 post- MRSA inoculation, mice were administered BFJD and linezolid by oral gavage intervention, and after 14 days of treatment, the mice were euthanized for sampling. (B) Bacterial burdens in the lungs, liver, and kidneys of mice (n=6). (C) The gating strategy of macrophages (CD11b+F4/80+) in mouse lung tissue flow cytometry. (D) The percentage of macrophages in the Ly6G - cell population and relative count in total harvested cells (n=5). (E) Representative FACS plots of macrophage polarization. (F) The percentage of M1- type (CD80+CD206-) macrophages and MFI of CD80 (n=5). (G) Representative immunofluorescence micrograph of lung tissue stained with indicated antibodies for CD68, CD86 and CD206. Nuclei were revealed by DAPI staining. Scale bars, 200 μm. Data are presented as the mean ± SD. Differences were analyzed applying ordinary one-way ANOVA followed by Dunnett´s multiple comparisons test (comparison with the Infected Model). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Subsequently, the lung tissue sections were incubated with primary antibodies against CD68 (BA3638, Boster, 1:200), CD86 (CY5238, ABways, 1:200) and CD206 (PTG, 60143-1-Ig, 1:200) overnight at 4°C.

Techniques: Infection, In Vivo, Sampling, Flow Cytometry, Immunofluorescence, Staining, Comparison

Journal: IBRO neuroscience reports

Article Title: IL-33 relieves nerve injury by mediating microglial polarization in neuromyelitis optica spectrum disorders via the IL-33/ST2 pathway.

doi: 10.1016/j.ibneur.2024.07.008

Figure Lengend Snippet: Fig. 5. Microglial cells were activated and transformed to the M2 phenotype after treatment with IL-33. (a) Western blot images and the quantification of CD206 and CD40 levels after treatment with IL-33 or NMOD serum. (b) BV2 cells were labeled with Iba-1 and CD68 for immunofluorescence. (c) Expression levels of PSD95 were observed by immunofluorescence after treatment with IL-33 or NMOSD serum. One-way analysis of variance was used for statistical analyses. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: For protein immunoreactivity in cells, the primary cortical neurons or BV2 cells were fixed with 4 % paraformaldehyde for 30 min at RT and treated with 0.5 % Triton for 10 min. After being washed with PBS, the cells were blocked with 5 % bovine serum albumin (BSA) for 1 h and then incubated with a rabbit antiPSD95 antibody (1:100 dilution, PA2295, Boster), anti-CD68 antibody (1:200 dilution, 28058–1-AP, Proteintech), anti-IBA1 antibody (1:100 dilution, CY7217, Abways), or anti-ST2 antibody (1:200 dilution, PRS3363, Sigma) at 4 ◦C overnight.

Techniques: Transformation Assay, Western Blot, Labeling, Immunofluorescence, Expressing

Journal: International Dental Journal

Article Title: Inhibition of Dynamin-Related Protein 1-Dependent Mitochondrial Fission Ameliorates Apical Periodontitis by Attenuating NLRP3 Inflammasome-Mediated M1 Macrophage Polarisation

doi: 10.1016/j.identj.2025.100853

Figure Lengend Snippet: Imbalance of mitochondrial dynamics mediated by p-Drp1 (Ser616) exists in macrophages from human AP samples. (A) Representative image of p-Drp1 (Ser616) IHC staining for the NC, RC, and PG group. Scale bar =50 μm. Higher magnification of the region of interest in cartilage is shown below each image. The quantification of percentage of p-Drp1 (Ser616) positive cells. (B) Representative western-blotting images of p-Drp1 (Ser616) and Drp1 and quantification in different groups. (C) Representative immunofluorescence images identified the colocalisation of p-Drp1 (Ser616) /TOMM20, MFN2/TOMM20. The yellow dotted line indicated the colocalisation between p-Drp1 (Ser616) -TOMM20 and MFN2-TOMM20. The quantification of percentage of p-Drp1 (Ser616) -TOMM20 and MFN2-TOMM20 double-positive cells in different groups. Scale bar = 50 μm. (D) Representative immunofluorescence images and quantifications of the ration of p-Drp1 (Ser616) /MFN2 in different groups. Scale bar = 50 μm. (E) Representative immunofluorescence images and quantifications of the expression of p-Drp1 (Ser616) and MFN2 in macrophages in different groups. Scale bar = 50 μm. (F) Pearson correlation between histoscore of p-Drp1 (Ser616) and histoscore of CD86, Nlrp3, Cleaved-caspase1 or IL-1β in periapical lesions. All data were analysed by one-way ANOVA and presented as mean ± SD (n=3). * P < .05, ** P < .01, *** P < .001.

Article Snippet: The sections were incubated with primary antibodies: anti-p-Drp1 (Ser616) (ABclonal), anti-NLRP3(Abways), anti-MFN2, anti-CD86 (Bioss), anti-Cleaved Caspase1(ZenBio) anti-CD68, anti-TOMM20 and anti-IL1-β (Santa Cruz) for 18 to 24 hours.

Techniques: Immunohistochemistry, Western Blot, Immunofluorescence, Expressing

Journal: International Dental Journal

Article Title: Inhibition of Dynamin-Related Protein 1-Dependent Mitochondrial Fission Ameliorates Apical Periodontitis by Attenuating NLRP3 Inflammasome-Mediated M1 Macrophage Polarisation

doi: 10.1016/j.identj.2025.100853

Figure Lengend Snippet: Pg-LPS induces abnormal mitochondrial division, activation of NLRP3 inflammatory pathway and M1 polarisation in macrophages. (A) Representative western-blotting images and quantification in different groups. (B) Representative images of ROS staining and quantification of intensity units in each group. Scale bar = 50 μm. (C) Representative images of JC-1 staining in different groups and quantification of red and green fluorescence intensity. Scale bar = 50 μm. (D) Quantification data of ATP level in macrophages. (E) Representative immunofluorescence images identified the colocalisation of p-Drp1 (Ser616) /TOMM20. Plots of pixel intensity of p-Drp1 (Ser616) with TOMM20. Scale bar = 10 μm. (F) Representative images of MitoTracker Red stain immunofluorescence (white boxes indicate higher magnification area of corresponding lower panels). Quantification of mitochondrial number and length. n = 25 Scale bar = 10 μm. (G) Mitochondrial morphology was visualised by transmission electron microscopy. Scale bar = 1 μm. All data were analysed by one-way ANOVA and presented as mean ± SD (n = 3). * P < .05, ** P < .01, *** P < .001.

Article Snippet: The sections were incubated with primary antibodies: anti-p-Drp1 (Ser616) (ABclonal), anti-NLRP3(Abways), anti-MFN2, anti-CD86 (Bioss), anti-Cleaved Caspase1(ZenBio) anti-CD68, anti-TOMM20 and anti-IL1-β (Santa Cruz) for 18 to 24 hours.

Techniques: Activation Assay, Western Blot, Staining, Fluorescence, Immunofluorescence, Transmission Assay, Electron Microscopy

Journal: International Dental Journal

Article Title: Inhibition of Dynamin-Related Protein 1-Dependent Mitochondrial Fission Ameliorates Apical Periodontitis by Attenuating NLRP3 Inflammasome-Mediated M1 Macrophage Polarisation

doi: 10.1016/j.identj.2025.100853

Figure Lengend Snippet: Mdivi-1 inhibits the activation of NLRP3 inflammatory pathway and M1 polarisation by inhibiting abnormal mitochondrial fission. (A) The protein levels of NLRP3 and CD86 were measured by Western blotting. Quantifications are on the right side. The protein levels of CD86 were measured by Western blotting. Quantifications are on the right side. (C) The protein levels of NLRP3, pro-caspase-1, caspase-1, pro-IL-lβ and IL-lβ were measured by Western blotting. Quantifications are on the right side. (D) Representative immunofluorescence images identified the colocalisation of p-Drp1 (Ser616) /TOMM20. Plots of pixel intensity of p-Drp1 (Ser616) with TOMM20. Scale bar = 10 μm. Quantifications are on the right side. (E) Mitochondrial morphology was visualised by transmission electron microscopy. Scale bar = 1 μm. (F) Representative images of ROS staining immunofluorescence (white boxes indicate higher magnification area of corresponding lower panels) and quantification of intensity units in each group. Scale bar = 50 μm. (G) Representative images of JC-1 staining in different groups and quantification of red and green fluorescence intensity. Scale bar = 50 μm. (H) Quantification data of ATP level in macrophages. All data were analysed by one-way ANOVA and presented as mean ± SD (n = 3). * P < .05, ** P < .01, *** P < .001.

Article Snippet: The sections were incubated with primary antibodies: anti-p-Drp1 (Ser616) (ABclonal), anti-NLRP3(Abways), anti-MFN2, anti-CD86 (Bioss), anti-Cleaved Caspase1(ZenBio) anti-CD68, anti-TOMM20 and anti-IL1-β (Santa Cruz) for 18 to 24 hours.

Techniques: Activation Assay, Western Blot, Immunofluorescence, Transmission Assay, Electron Microscopy, Staining, Fluorescence

Journal: International Dental Journal

Article Title: Inhibition of Dynamin-Related Protein 1-Dependent Mitochondrial Fission Ameliorates Apical Periodontitis by Attenuating NLRP3 Inflammasome-Mediated M1 Macrophage Polarisation

doi: 10.1016/j.identj.2025.100853

Figure Lengend Snippet: Mdivi-1 can inhibit periapical lesions in experimental periapical periodontitis mice. (A) Flow diagram of the treatment of apical periodontitis (AP) with Mdivi-1 in C57BL/6 mice. (B) Schematic diagram of access opening for experimental AP. (C) The body weight of mice in different groups. (D) The representative micro-CT images of experimental AP tissues of the mice. The images were taken using 3-dimensional micro-Ct at 3 different views: coronal, sagittal and horizontal views. Quantification of bone loss volume are on the right side. (E) Representative images of HE staining and TRAP staining of experiment AP in different group. Quantification of TRAP staining are on the right side. Scale bar = 100 μm. (F) Representative image of CD86, NLRP3, Cleaved-Caspase1, IL1β and p-Drp1 (Ser616) immunohistochemistry (IHC) staining for the different groups, hematoxylin indicates cell nucleus. Scale bar = 100 μm. Higher magnification of the region of interest in cartilage is shown below each image. The quantification of percentage of CD86, NLRP3, Cleaved-Caspase1, IL1β and p-Drp1 (Ser616) positive cells. (G) Representative immunofluorescence images and quantifications of the expression of CD86, NLRP3, Cleaved-Casepase1 and p-Drp1 (Ser616) with F4/80 in different groups. Scale bar = 100 μm. All data were analysed by one-way ANOVA and presented as mean ± SD (n = 3). * P < .05, ** P < .01, *** P < .001.

Article Snippet: The sections were incubated with primary antibodies: anti-p-Drp1 (Ser616) (ABclonal), anti-NLRP3(Abways), anti-MFN2, anti-CD86 (Bioss), anti-Cleaved Caspase1(ZenBio) anti-CD68, anti-TOMM20 and anti-IL1-β (Santa Cruz) for 18 to 24 hours.

Techniques: Micro-CT, Staining, Immunohistochemistry, Immunofluorescence, Expressing

Journal: International Dental Journal

Article Title: Inhibition of Dynamin-Related Protein 1-Dependent Mitochondrial Fission Ameliorates Apical Periodontitis by Attenuating NLRP3 Inflammasome-Mediated M1 Macrophage Polarisation

doi: 10.1016/j.identj.2025.100853

Figure Lengend Snippet: Schematic diagram illustrating the p-Drp1 (Ser616) -dependent mitochondrial fission involved in the sophisticated signaling pathways of apical periodontitis.

Article Snippet: The sections were incubated with primary antibodies: anti-p-Drp1 (Ser616) (ABclonal), anti-NLRP3(Abways), anti-MFN2, anti-CD86 (Bioss), anti-Cleaved Caspase1(ZenBio) anti-CD68, anti-TOMM20 and anti-IL1-β (Santa Cruz) for 18 to 24 hours.

Techniques: Protein-Protein interactions

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