mouse mononuclear macrophage leukemia cell line raw264 7  (ATCC)

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: Cell Reports Medicine

Article Title: Restoring immune homeostasis in atherosclerotic plaques via inorganic violet phosphorus nano-immunotherapy

doi: 10.1016/j.xcrm.2025.102528

Figure Lengend Snippet: Preparation, characterization, ROS scavenging, and biocompatibility of VPNS@P (A) Schematic of exfoliation and PEGylation of violet phosphorus (VP) to prepare VPNS@P from bulk VP. (B) Transmission electron microscopy (TEM) image of VPNS@P. Scale bar, 100 nm. (C) Hydrodynamic size distribution of VPNS@P measured by dynamic light scattering (DLS). (D and E) Atomic force microscopy (AFM) image (D) and thickness profile (E) of VPNS@P. Scale bar, 100 nm. (F) Raman scattering spectra of VPNS@P. (G) Time-course DLS measurements of VPNS@P incubated in PBS or DMEM supplemented with 10% fetal bovine serum (FBS) over 7 days ( n = 3 independent samples). (H–J) Scavenging capability of VPNS@P ( n = 5 independent samples) toward H 2 O 2 (H), ·OH (I), and O 2 ·− (J). (K–M) Biocompatibility of VPNS@P in vitro . Cell viabilities of RAW264.7 (K), mouse aortic vascular smooth muscle cells (MOVASs) (L), and human umbilical vein endothelial cells (HUVECs) (M) were examined with a CCK-8 assay ( n = 3 biologically independent samples). Data were analyzed using one-way ANOVA with a Dunnett’s T3 post hoc test and are shown as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. ns, not significant.

Article Snippet: Mouse: RAW264.7 macrophage cell line , ATCC , TIB-71; RRID: CVCL_0493.

Techniques: Transmission Assay, Electron Microscopy, Microscopy, Incubation, In Vitro, CCK-8 Assay