raw 264 7 murine macrophage cells  (ATCC)

Journal: Bioactive Materials

Article Title: Bioengineered titanium implants functionalized with aptamer-valproic acid conjugates orchestrate macrophage programming and mesenchymal stem cell homing for improved osseointegration

doi: 10.1016/j.bioactmat.2026.05.055

Figure Lengend Snippet: A) Live/Dead staining of RAW 264.7 macrophages cultured on different material surfaces for 72 h, where dead cells were stained red and live cells were stained green. B) Cytoskeleton staining morphology of RAW 264.7 grown on different material surfaces for 72 h. C, D, F) Fluorescence images and flow cytometry analysis of intracellular ROS in RAW 264.7 cells with DCFH-DA probe. E) Proliferation of RAW 264.7 macrophages on different material surfaces assessed by CCK-8 assay. n = 3. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Created in BioRender.

Article Snippet: The murine macrophage cell line RAW 264.7 (Procell Life Science, China) was cultured in high-glucose DMEM (Gibco, USA) containing 10% FBS and employed for immunomodulation studies.

Techniques: Staining, Cell Culture, Fluorescence, Flow Cytometry, CCK-8 Assay

Journal: Bioactive Materials

Article Title: Bioengineered titanium implants functionalized with aptamer-valproic acid conjugates orchestrate macrophage programming and mesenchymal stem cell homing for improved osseointegration

doi: 10.1016/j.bioactmat.2026.05.055

Figure Lengend Snippet: A, F) IF staining of iNOS and CD206 in RAW 264.7 macrophages. B-E) Secretion of inflammation-related proteins in RAW 264.7 macrophages. G-J) Relative mRNA expression of inflammation-related genes in RAW 264.7 macrophages. n = 3. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The murine macrophage cell line RAW 264.7 (Procell Life Science, China) was cultured in high-glucose DMEM (Gibco, USA) containing 10% FBS and employed for immunomodulation studies.

Techniques: Staining, Expressing

Journal: RSC Advances

Article Title: Salmon bone-derived chondroitin sulfate nanogels for controlled delivery of curcumin: release behavior, structural characteristics and anti-inflammatory efficacy

doi: 10.1039/d6ra06378k

Figure Lengend Snippet: (a) Inhibitory effects on nitric oxide (NO) production and (b) cell viability of RAW 264.7 macrophages treated with blank CS, free curcumin, physical mixture of CS + curcumin (Mix CS + Cur), and CS-Cur nanogel. Data represent mean ± SD ( n = 3, independent replicates). Statistical significance in (a) relative to the curcumin control group is denoted as: * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Article Snippet: For the in vitro cell culture and biological evaluation, the RAW 264.7 murine macrophage cell line was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Control

Journal: Frontiers in Immunology

Article Title: Targeting the capsule of Klebsiella pneumoniae with a cationic CR3-binding protein enhances phagocytosis and promotes bacterial clearance and survival in a mouse sepsis model

doi: 10.3389/fimmu.2026.1934439

Figure Lengend Snippet: rPF4 enhances phagocytosis of heat-inactivated and live low-virulence K. pneumoniae by macrophages. (A) Effect of rPF4 on phagocytosis of K. pneumoniae by suspended IC-21 macrophages. Cells were incubated with fluorescein-labeled, heat-inactivated K. pneumoniae (ATCC 13883; capsule type K3) pretreated with 50 µg/ml rPF4 for 30 minutes at 37 °C. After quenching fluorescence with ethidium bromide, phagocytosis was quantified by flow cytometry. Representative histograms from eight independent experiments are shown. (B) Quantification of the effect of different concentrations of rPF4 on phagocytosis of K. pneumoniae by suspended mouse IC-21 macrophages based on flow cytometry results. MFI, mean fluorescence intensity. Data are presented as the mean fold difference in MFI ± standard deviation from 3–8 independent experiments. *p < 0.05, ** p < 0.01, *** p < 0.001, ns, nonsignificant. (C–F) Quantification of phagocytosis of K. pneumoniae by various phagocytes in suspension in the presence of different concentrations of rPF4. Values represent MFI fold-difference means ± S.D. from 3–8 independent experiments. (G) Fluorescein-labeled, heat-inactivated K. pneumoniae were pretreated with 50 µg/ml rPF4 and added to adherent TG-elicited inflammatory macrophages for 60 minutes at 37 °C. Non-phagocytosed bacteria were removed, and phagocytosis was assessed from fluorescent cell images. Representative images of macrophages exposed to rPF4-coated bacteria ( upper panel ) and uncoated control bacteria ( lower panel ) are shown. The scale bars are 10 μm. (H) Representative confocal image showing phagocytosed fluorescein-labeled K. pneumoniae inside a macrophage. Representative confocal image showing phagocytosed fluorescein-labeled K. pneumoniae inside a macrophage with orthogonal YZ and XZ views. The scale bar is 5 μm. The scale bar is 5 μm. (I) Quantification of rPF4-mediated enhancement of K. pneumoniae phagocytosis, expressed as the number of fluorescent bacteria per cell. Phagocytosis was assessed in IC-21 macrophages, mouse inflammatory macrophages, and mouse neutrophil/monocyte populations. Values represent mean ± S.D. from three random fields per condition in three independent experiments ***p < 0.001. (K) Live pHrodo-labeled K. pneumoniae were preincubated with rPF4 and then added to various macrophages in suspension. After 30 min at 37 °C, phagocytosis was measured by flow cytometry. Values represent mean fold differences in MFI ± S.D. from 4–9 independent experiments; ***p < 0.001. (L) Adherent IC-21 macrophages (5×10 5 ) were incubated with 10 6 CFU of K. pneumoniae pretreated with 50 μg/ml rPF4 for 1 h at 37 °C. After phagocytosis, cells were washed, and extracellular bacteria that may have remained on the cell surface were inactivated with gentamicin for 1 h. Macrophages were then either lysed immediately with 0.2% Triton X-100 or cultured in DMEM for 3 and 24 hours. Aliquots of cell lysates were plated on LB agar plates for 16 h at 37 °C, and CFUs were determined. Data are means ± S.D. from 3 experiments. ***p< 0.001.

Article Snippet: The IC-21 murine macrophage cell line, human monocytic U937 and THP-1 cells, and A549 lung epithelial cells were obtained from ATCC and cultured in RPMI supplemented with 10% FBS and antibiotics.

Techniques: Incubation, Labeling, Fluorescence, Flow Cytometry, Standard Deviation, Suspension, Bacteria, Control, Confocal, Cell Culture

Journal: Frontiers in Immunology

Article Title: Targeting the capsule of Klebsiella pneumoniae with a cationic CR3-binding protein enhances phagocytosis and promotes bacterial clearance and survival in a mouse sepsis model

doi: 10.3389/fimmu.2026.1934439

Figure Lengend Snippet: rPF4 enhances phagocytosis of heat-inactivated and live high-virulence K. pneumoniae . (A–D) Heat-inactivated pHrodo-labeled hv Kp (ATCC 43816; capsule type K2) were preincubated with various concentrations of rPF4 for 1 hour at 37 °C and rPF4-coated bacteria were then added to suspended IC-21 macrophages, PMA-activated THP-1 cells, TG-elicited inflammatory mouse macrophages, and mouse peritoneal neutrophils/monocytes for 30 minutes at 37 °C, and phagocytosis was measured by flow cytometry. (E–H) , Live bacteria were labeled with pHrodo, preincubated with rPF4, and then added to various leukocyte populations to assess phagocytosis. Data are presented as means ± S.D. from three separate experiments. ***p< 0.001. (I) The table shows fold changes in phagocytosis at two concentrations of rPF4 (50 and 100 μg/ml) relative to phagocytosis in the absence of rPF4 across various macrophage types.

Article Snippet: The IC-21 murine macrophage cell line, human monocytic U937 and THP-1 cells, and A549 lung epithelial cells were obtained from ATCC and cultured in RPMI supplemented with 10% FBS and antibiotics.

Techniques: Labeling, Bacteria, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Isobavachalcone attenuates experimental periodontitis by suppressing PTGS2-associated macrophage ferroptosis and restoring periodontal immune homeostasis

doi: 10.3389/fphar.2026.1913548

Figure Lengend Snippet: IBC mitigates macrophage ferroptosis via the PTGS2-MAPK-ROS-GPX4 axis. (A–G) Representative Western blots and quantitative analysis of iNOS, PTGS2, p-JNK, JNK, p-p38, p38, GPX4, and IL-1β in LPS-stimulated RAW264.7 cells treated with IBC in a dose-dependent manner. (H, J) Detection and quantification of iNOS, PTGS2, MAPK signaling proteins, and GPX4 in gingival tissues collected from rat models of experimental periodontitis. (I, K) Representative Western blots and quantitative analysis of the PTGS2-MAPK-GPX4 signaling axis in macrophages following OE-PTGS2 plasmid transfection and IBC treatment to assess pathway rescue. (L–P) Microscopic visualization of MitoSOX and FerroOrange signals, accompanied by the measurement of fluorescence intensity and MDA levels in cells undergoing OE-PTGS2 rescue treatments. (Q–T) Representative C11-BODIPY fluorescence imaging, GSH/GSSG ratio, and MDA content assessment in macrophages treated with the ROS scavenger NAC. Data Note: Data are presented as mean ± SD from three independent biological replicates (n = 6) for in vitro experiments, and n = 6 per group for in vivo analysis. Scale bar = 50 μm.

Article Snippet: The RAW264.7 murine macrophage cell line was obtained from the American Type Culture Collection (ATCC, Manassas, VA, United States of America).

Techniques: Western Blot, Analysis, Plasmid Preparation, Transfection, Fluorescence, Imaging, In Vitro, In Vivo