Journal: Journal of Advanced Research

Article Title: PEBL, a component-based Chinese medicine, reduces virus-induced acute lung injury by targeting FXR to decrease ACE2 levels

doi: 10.1016/j.jare.2025.05.003

Figure Lengend Snippet: PEBL alleviates Poly(I:C)-induced ALI in a dose-dependent manner and modulates cytokine levels in macrophage inflammation. (A) Experimental design for PEBL treatment in ALI zebrafish. (B) Dose-dependent reduction in mortality by PEBL. Survival plot of 5 dpf Tg(coro1α: GFP) larvae at 72 hpi ( n = 30). (C) Dose-dependent reduction in macrophage recruitment by PEBL. Quantitative analysis of macrophage infiltration in the swim bladder section at 4 hpi ( n = 10). (D) Fluorescence images of macrophages in the swim bladder section at 4 hpi following different concentrations of PEBL, marked by the red circle. (E-J) PEBL reduces Poly(I:C)-induced cytokine elevation in RAW264.7 cells ( n = 3). mRNA levels of IL-1β, IL-6, and TNF-α in cells were measured by qPCR (E-G), while protein concentrations of these cytokines in culture media were quantified using ELISA (H-J). ## P < 0.01, ### P < 0.001 vs. Poly(I:C); ** P < 0.01, *** P < 0.001 for group comparisons. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Mouse peritoneal mononuclear macrophage RAW264.7 cells and human embryonic kidney 293 T cells were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: Journal of Advanced Research

Article Title: PEBL, a component-based Chinese medicine, reduces virus-induced acute lung injury by targeting FXR to decrease ACE2 levels

doi: 10.1016/j.jare.2025.05.003

Figure Lengend Snippet: PEBL suppresses Poly(I:C)-induced FXR and ACE2 expression and NF-κB-p65 nuclear translocation in RAW264.7 cells. (A-E) PEBL reduces the mRNA (A-B) and protein (D-E) levels of FXR and ACE2 and diminishes NF-κB-p65 nuclear translocation (C, E). (F-H) PEBL suppresses the protein distribution of FXR and ACE2, inhibits the nuclear translocation of NF-κB-p65. Representative images show the localization of FXR (F, green), ACE2 (G, green), NF-κB-p65 (H, green), and DAPI (blue), captured by immunofluorescence at 40 × magnification using confocal microscopy. Scale bar = 10 μm. UDCA was used as a positive control. Nuc, nucleus; Cyt, cytoplasm; Mem, membrane. n = 3; * P < 0.05, ** P < 0.01, *** P < 0.001 for group comparisons. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Mouse peritoneal mononuclear macrophage RAW264.7 cells and human embryonic kidney 293 T cells were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Expressing, Translocation Assay, Immunofluorescence, Confocal Microscopy, Positive Control, Membrane

Journal: Journal of Advanced Research

Article Title: PEBL, a component-based Chinese medicine, reduces virus-induced acute lung injury by targeting FXR to decrease ACE2 levels

doi: 10.1016/j.jare.2025.05.003

Figure Lengend Snippet: PEBL suppresses Poly(I:C)-induced FXR binding to ACE2 by inhibiting FXR transcription in RAW264.7 cells. (A-B) FXR overexpression reverses the effect of PEBL on the protein levels of ACE2 and NF-κB-p65. n = 3. (C-D) FXR overexpression reverses the inhibitory effect of PEBL on ACE2 distribution and NF-κB-p65 nuclear translocation. Representative images show the localization of ACE2 (C, green), NF-κB-p65 (D, green), and DAPI (blue), captured by immunofluorescence at 40 × magnification using confocal microscopy. Scale bar = 10 μm. (E-H) PEBL requires FXR to decrease ACE2 expression and mitigate Poly(I:C) infection. In FXR-KD cells (F, H), no significant change in ACE2 expression was observed following treatments with CDCA, Poly(I:C), UDCA, or PEBL, compared to WT cells (E, G). WT, wild-type RAW264.7 cells; n = 3. (I) Co-IP analysis reveals no binding between FXR and ACE2 proteins. (J-K) PEBL reduces Poly(I:C)-induced FXR binding to the ACE2 promoter, confirmed by ChIP-qPCR and agarose gel electrophoresis.Nuc, nucleus; Cyt, cytoplasm; Mem, membrane; OSTα, positive control; ACE2-NC, negative control; C, control; P, Poly(I:C). n = 6; * P < 0.05, ** P < 0.01, *** P < 0.001 for group comparisons; ns , non-significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Mouse peritoneal mononuclear macrophage RAW264.7 cells and human embryonic kidney 293 T cells were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Binding Assay, Over Expression, Translocation Assay, Immunofluorescence, Confocal Microscopy, Expressing, Infection, Co-Immunoprecipitation Assay, ChIP-qPCR, Agarose Gel Electrophoresis, Membrane, Positive Control, Negative Control, Control

Journal: bioRxiv

Article Title: GW4869 depletes macrophages and increases number of extracellular vesicles in murine peritoneal cavity fluid

doi: 10.64898/2026.01.20.700528

Figure Lengend Snippet: GW4869 depletes peritoneal macrophages and induces local inflammation. A) Schematic overview of treatment schedule. B) Total peritoneal cells per mouse (p = 0.00151). C) Viability of cells (p = 0.00154). D) Stacked bar charts of cell types represented as frequency of CD45 + cells. E) Bar charts of total number of macrophages (p = <0.0001), monocytes (p = 0.0385), neutrophils (p = 0.0017, p = 0.0282) and eosinophils (p = 0.036). F) Representative bivariate plots of LPM and SPM populations. G) Bar chart showing total LPMs (p = <0.0001) and SPMs (p = <0.0001) per mouse. H) CD11b MFI of neutrophils (p = <0.0001) and eosinophils (p = <0.0001). n = 7.

Article Snippet: Peritoneal cells were counted on a NucleoCounter NC-3000 using A8 NC-slides and viability solution 13 (ChemoMetec).

Techniques:

Journal: bioRxiv

Article Title: GW4869 depletes macrophages and increases number of extracellular vesicles in murine peritoneal cavity fluid

doi: 10.64898/2026.01.20.700528

Figure Lengend Snippet: GW4869 increases number of total peritoneal EVs. A) Brief schematic overview of sample workflow. B) Gating strategy for nano flow cytometry analyses peritoneal particles and EVs. C) Bar chart of total particles per mouse derived from inclusion gate (p = 0.00142). D) Total EVs per mouse (p = 0.00338). E) Total CD9 + EVs per mouse (p = 0.01146). F) Total CD9 + cells per mouse (p = 0.00026). G) Median size in nm derived from size calibration using FCMPASS within inclusion gate, EV gate (p = <0.0001, p = 0.00062), and CD9 + EVs. n = 7.

Article Snippet: Peritoneal cells were counted on a NucleoCounter NC-3000 using A8 NC-slides and viability solution 13 (ChemoMetec).

Techniques: Flow Cytometry, Derivative Assay

Journal: bioRxiv

Article Title: GW4869 depletes macrophages and increases number of extracellular vesicles in murine peritoneal cavity fluid

doi: 10.64898/2026.01.20.700528

Figure Lengend Snippet: Repeat experiment of GW4869 i.p. 24 h time point. A) Total peritoneal cells per mouse (p = 0.0102). B) Viability of cells. C) Bar chart of total particles per mouse derived from inclusion gate. D) Total EVs per mouse (p = 0.0028). E) Total CD9 + EVs per mouse (p = 0.0005). n = 8.

Article Snippet: Peritoneal cells were counted on a NucleoCounter NC-3000 using A8 NC-slides and viability solution 13 (ChemoMetec).

Techniques: Derivative Assay

Journal: bioRxiv

Article Title: GW4869 depletes macrophages and increases number of extracellular vesicles in murine peritoneal cavity fluid

doi: 10.64898/2026.01.20.700528

Figure Lengend Snippet: Macrophage depletion increases number of total peritoneal EVs. A) Illustration of mouse model and treatment applied. B) Total peritoneal cells per mouse (p = 0.0332). C) Viability of cells. D) Bar chart showing macrophage frequency of CD45 + cells (p = 0.0009). E) Stacked bar charts of cell types represented as frequency of CD45 + cells and as total cells per mouse. F) Bar charts representing total macrophages (p = 0.0096), monocytes (p = 0.0013), neutrophils (p = 0.0031), and eosinophils per mouse. G) Bar chart of total particles per mouse derived from inclusion gate. H) Total EVs per mouse (p = 0.005). I) Total CD9 + EVs per mouse (p = 0.0048). n = 4.

Article Snippet: Peritoneal cells were counted on a NucleoCounter NC-3000 using A8 NC-slides and viability solution 13 (ChemoMetec).

Techniques: Derivative Assay

Journal: PLOS Pathogens

Article Title: Mycobacterium tuberculosis triggers reduced inflammatory cytokine responses and virulence in mice lacking Tax1bp1

doi: 10.1371/journal.ppat.1012829

Figure Lengend Snippet: (A and B) BMDMs or peritoneal exudate cells were infected with L. monocytogenes, and CFU were counted at 30 minutes, 2-, 5-, or 8 hours post-infection. Results are the mean ± SEM from three technical replicate samples. The p values from t-test comparisons are shown. (C and D) CFU from spleen and liver homogenates from mice intravenously infected with L. monocytogenes were enumerated at 4-, 10-, or 48-hours post-infection. Results are the mean ± SEM from five mice. Brackets indicate p values from t-test comparisons. (E & G) Cytokine levels were measured from the serum of mice infected with L. monocytogenes at 4-, 10-, and 48-hours post-infection by cytometric bead array (IL-6, TNF-α, IFN-γ, MCP-1, IL-10) or ELISA (IFN-β). Results are mean ± SEM from five samples. Brackets indicate p values from t - test comparisons. (F) Spleen and liver homogenates from mice intraperitoneally infected with L. monocytogenes were enumerated for CFU 72 hours post-infection. Results are the mean ± SEM from five mice. Brackets indicate p values from t-test comparisons. CFU data were logarithmically transformed prior to statistical analysis.

Article Snippet: Biological sample ( M. musculus ) , Primary bone marrow-derived macrophages, AMs, and peritoneal exudate cells, wild-type C57BL/6J , Jackson Laboratory , Stock # 000664 , .

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Transformation Assay