raw264 7  (ATCC)

Journal: Bioactive Materials

Article Title: A foam cell-targeted lipophagy restoration strategy stabilizes vulnerable atherosclerotic plaques

doi: 10.1016/j.bioactmat.2026.02.041

Figure Lengend Snippet: In vitro evaluation of foam cell lipid accumulation and lipophagy activation following OPN-HMCN@MLT treatment. ( A - C ) ORO and BODIPY staining images and corresponding quantification of ORO and BODIPY positive areas of RAW264.7 cells under different stimulations (n = 5, scale bar for ORO: 100 μm, scale bar for BODIPY: 20 μm). ( D ) Bio-TEM images of RAW264.7 cells post various treatments (n = 5, scale bars 1.0 μm). Green arrows indicate nanoparticles. ( E , F ) Morphometric analysis determined the mean number and area (μm 2 ) of LDs per cell section. ( G ) Confocal images depicting lipophagy flux in foam cells following different treatments (n = 5 biological replicates, scale bars: 10 μm). ( H - J ) The quantities of acidified autophagosomes (GFP-RFP+), neutral autophagosomes (GFP + RFP+), and LDs labeled with BODIPY were measured per cell for each condition. (K to N) Representative Western blot images and quantitative analysis of LC3, LAMP1, and P62 expression in foam cells. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: RAW264.7 mouse macrophage cells (ATCC® TIB-71; RRID: CVCL_0493) and MCAECs (Procell, CP-M081) were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% fetal bovine serum and 1% penicillin-streptomycin at 37 °C in a 5% CO 2 atmosphere.

Techniques: In Vitro, Activation Assay, Staining, Labeling, Western Blot, Expressing

Journal: Bioactive Materials

Article Title: A foam cell-targeted lipophagy restoration strategy stabilizes vulnerable atherosclerotic plaques

doi: 10.1016/j.bioactmat.2026.02.041

Figure Lengend Snippet: In vitro examination of LD degradation in foam cells through fatty acid oxidation and cholesterol efflux. (A ) Schematic diagram of the LD degradation mechanism. ( B , C ) Confocal images and quantitative analysis of LDs colocalization with fatty acids in RAW264.7 cells following different treatments (n = 5, scale bars: 5 μm). ( D , E ) Confocal images of the colocalization of mitochondria with fatty acids and quantified data of fatty acids in RAW264.7 cells under different stimulations (n = 5, scale bars: 5 μm). ( F , G ) Confocal images illustrating mitochondrial colocalization with ATP and corresponding quantification of ATP levels in RAW264.7 cells post various treatments (n = 5, scale bars: 20 μm). ( H ) Diagram illustrating the incorporation of [U- 13 C] palmitic acid into the TCA cycle and the labeling pattern of derived metabolites (n = 3). ( I ) A PCA plot illustrates the cluster separation between the two groups (n = 3). ( J ) Heatmap showing differences in metabolites between the two groups (n = 3). ( K ) Normalized total labeling of each metabolite to [U- 13 C] palmitic acid (n = 3). ( L ) Proportion of (m + 2)-labeled TCA cycle metabolites derived from [U- 13 C] palmitic acid (n = 3). ( M - R ) The study quantified NBD-cholesterol accumulation ( M , P ) and cholesterol efflux facilitated by HDL ( N , O ) and apoA-I ( Q , R ) using confocal imaging across (n = 5, Scale bar = 50 μm). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: RAW264.7 mouse macrophage cells (ATCC® TIB-71; RRID: CVCL_0493) and MCAECs (Procell, CP-M081) were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% fetal bovine serum and 1% penicillin-streptomycin at 37 °C in a 5% CO 2 atmosphere.

Techniques: In Vitro, Labeling, Derivative Assay, Imaging

Journal: Journal of Cell Science

Article Title: Selective disruption of microtubule formation at the nuclear envelope impairs the bone resorption capacity of osteoclasts

doi: 10.1242/jcs.264166

Figure Lengend Snippet: MTOC proteins and Golgi are recruited to the nuclear envelope in RAW264.7-derived osteoclasts. (A) Immunostaining of AKAP6 (green) and GM130 (magenta) together with DNA (DAPI) in RAW264.7-derived osteoclasts after 4 days of RANKL addition. Scale bars: 10 µm. Immunostaining of (B) PCNT (green), (C) CDK5RAP2 (green) and (D) PCM1 (green) and γ-tubulin (magenta) together with DNA (DAPI) in RAW264.7-derived osteoclasts after 4 days of RANKL addition. Arrows indicate centrosomes. Scale bars: 10 µm. Images representative of three experimental repeats.

Article Snippet: RAW264.7 cells were obtained from ATCC and maintained in high glucose DMEM supplemented with GlutaMAX (Thermo Fisher Scientific, 61965059) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco, 26140079), 1 mM sodium pyruvate, 100 U/ml penicillin and 100 μg/ml streptomycin (Thermo Fisher Scientific, 15140-122) and cultured at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Derivative Assay, Immunostaining

Journal: Journal of Cell Science

Article Title: Selective disruption of microtubule formation at the nuclear envelope impairs the bone resorption capacity of osteoclasts

doi: 10.1242/jcs.264166

Figure Lengend Snippet: RAW264.7-derived osteoclasts exhibit a nuclear envelope MTOC alongside a functional centrosome. (A) Immunostaining of α-tubulin (red), AKAP6 (green), GM130 (magenta) and DNA (DAPI) in RAW264.7-derived osteoclasts differentiated for 4 days. (B) Immunostaining of EB-1 (green), γ-tubulin (magenta) and DNA (DAPI) in RAW264.7-derived osteoclasts treated with nocodazole (upper row) and after 1 min (middle row) or 2 min (bottom row) recovery at 37°C. In A and B, asterisks indicate the perinuclear microtubule cage and arrows denote centrosomal asters. (C) Immunostaining of α-tubulin (red), PCNT (green), GM130 (magenta), and DNA (DAPI) in 4 day RAW264.7-derived osteoclasts after 1.5 min of recovery from nocodazole-induced microtubule depolymerization. Asterisks denote the NE nucleation, arrows denote centrosomal nucleation and arrowheads denote Golgi nucleation. Scale bars: 10 µm. Images representative of three experimental repeats.

Article Snippet: RAW264.7 cells were obtained from ATCC and maintained in high glucose DMEM supplemented with GlutaMAX (Thermo Fisher Scientific, 61965059) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco, 26140079), 1 mM sodium pyruvate, 100 U/ml penicillin and 100 μg/ml streptomycin (Thermo Fisher Scientific, 15140-122) and cultured at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Derivative Assay, Functional Assay, Immunostaining

Journal: Journal of Cell Science

Article Title: Selective disruption of microtubule formation at the nuclear envelope impairs the bone resorption capacity of osteoclasts

doi: 10.1242/jcs.264166

Figure Lengend Snippet: Nesprin-1α and AKAP6β isoforms predominate in RAW264.7-derived osteoclasts and form a functional LINC complex. (A) Immunostaining of AKAP6 (green), nesprin-1 (magenta) and DNA (DAPI) in RAW264.7 cells either non-differentiated (upper panels) or differentiated into osteoclasts for 4 days with 50 ng ml −1 of RANKL (lower panels). Non-differentiated cells lack detectable expression of both AKAP6 and nesprin-1 (also denoted with an arrow). Scale bars: 10 µm. (B) Heatmap of qPCR-derived RNA expression levels for Emr1 , Akap6 , Syne1a , Trap and Ctsk during osteoclast differentiation (days 1–4 of RANKL treatment) relative to non-differentiated precursor cells ( n =3). Expression values are normalized and represented as a percentage of maximal expression per gene using the viridis color map. (C) qPCR analysis of Syne1a (gray circles) and Syne1 giant (black triangles) mRNA expression during osteoclast differentiation (D1, D2, D3, D4, day 1–4). (D) qPCR analysis of Akap6a (gray circles) and Akap6b (black triangles) transcript levels at the indicated differentiation stages. Data in C and D are presented as mean±s.d. from three independent experiments, normalized to the non-differentiated condition. (E) Representative images of RAW264.7-derived osteoclasts treated with vehicle (water) or 5 μM DTT and co-stained with Sun2C (green), laminB1 (magenta) antibodies and DAPI. The Sun2C antibody recognizes an epitope which is masked when nesprin-1α is upregulated during osteoclast differentiation. DTT treatment restores epitope accessibility. Non-differentiated cells, indicated with an arrow, exhibit Sun2C reactivity. Scale bars: 10 µm. Images in A and E representative of three experimental repeats.

Article Snippet: RAW264.7 cells were obtained from ATCC and maintained in high glucose DMEM supplemented with GlutaMAX (Thermo Fisher Scientific, 61965059) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco, 26140079), 1 mM sodium pyruvate, 100 U/ml penicillin and 100 μg/ml streptomycin (Thermo Fisher Scientific, 15140-122) and cultured at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Derivative Assay, Functional Assay, Immunostaining, Expressing, RNA Expression, Staining

Journal: Journal of Cell Science

Article Title: Selective disruption of microtubule formation at the nuclear envelope impairs the bone resorption capacity of osteoclasts

doi: 10.1242/jcs.264166

Figure Lengend Snippet: AKAP6 knockdown disrupts NE-MTOC formation in RAW-derived osteoclasts. (A) qPCR analysis of mRNA expression in siControl- and siAKAP6-treated osteoclasts. Bars represent mean fold change (normalised to siControl) for the indicated genes, with individual experimental values shown as dots (three independent experiments). Akap6 expression was significantly reduced following AKAP6 depletion, whereas Syne1a , Syne1 giant , the macrophage marker Emr1 , and the osteoclast markers Ctsk , Nfatc1 and Trap transcript levels were not significantly affected. ***P <0.001; ns, not significant (two-way ANOVA followed by Bonferroni's multiple comparisons test). (B–F) Immunostaining in RAW264.7-derived osteoclasts transfected with control siRNA (siControl) or AKAP6 siRNA (siAKAP6) of (B) AKAP6 (green) and GM130 (magenta), and DNA (DAPI); (C) nesprin-1 (green), CDK5RAP2 (magenta), and DNA (DAPI); (D) PCM1 (green), GM130 (magenta), and DNA (DAPI); (E) PCNT, (green), γ-tubulin (magenta), and DNA (DAPI); and (F) CDK5RAP2 (green), γ-tubulin (magenta), and DNA (DAPI). Note that although perinuclear localization of PCNT and CDK5RAP2 is lost upon AKAP6 knockdown, centrosomal localization is unaffected (arrows). Scale bars: 10 μm. Images in B–F representative of three experimental repeats.

Article Snippet: RAW264.7 cells were obtained from ATCC and maintained in high glucose DMEM supplemented with GlutaMAX (Thermo Fisher Scientific, 61965059) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco, 26140079), 1 mM sodium pyruvate, 100 U/ml penicillin and 100 μg/ml streptomycin (Thermo Fisher Scientific, 15140-122) and cultured at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Knockdown, Derivative Assay, Expressing, Marker, Immunostaining, Transfection, Control

Journal: Journal of Cell Science

Article Title: Selective disruption of microtubule formation at the nuclear envelope impairs the bone resorption capacity of osteoclasts

doi: 10.1242/jcs.264166

Figure Lengend Snippet: AKAP6 Depletion disrupts NE-MTOC function, but not centrosomal-MTOC. (A) Immunostaining of α-tubulin (red), PCNT (green), GM130 (magenta) and DNA (DAPI) in RAW264.7-derived osteoclasts transfected with control siRNA (siControl) or AKAP6-targeting siRNA (siAKAP6). All stainings were performed in cells subjected to nocodazole washout (1.5 min). The yellow arrow indicates centrosomal microtubule organization and the arrowhead indicates Golgi-derived nucleation. (B) Quantification of fluorescence intensity profiles for α-tubulin, PCNT and GM130 in 0.2 µm wide concentric bands relative to distance from the nuclear edge (0.0). For each biological replicate ( n =3), 33–36 nuclei were analyzed per condition, and graphs show the mean±s.d. of the three independent biological experiments. The outward shift in the α-tubulin peak position is indicated by the red bracket, and the reduction in peak intensity is indicated by the green bracket. Statistical comparisons of peak position and peak intensity (amplitude) were performed at the experiment level with an unpaired two-tailed t -test; corresponding P -values are shown in the graphs. (C) Immunostaining of α-tubulin (red), EB1 (green), γ-tubulin (magenta) and DNA (DAPI) in RAW264.7-derived osteoclasts transfected with siControl or siAKAP6 performed after 1 min of nocodazole washout. Insets (2.5× magnification) highlight γ-tubulin-positive centrosomes exhibiting normal centrosomal microtubule outgrowth in both conditions. (D) Quantification of tubulin intensity in a 3 μm area surrounding each centrosome in siControl cells and siAKAP6 centrosomes from three independent experiments. SuperPlots display the overall mean (black line) and biological-replicate mean (color-coded triangles) from n =3, with the individual centrosomes (30–35 per condition) shown as matching color-coded dots superimposed beneath them. ns, not significant (paired two-tailed t -test). Scale bars: 10 μm.

Article Snippet: RAW264.7 cells were obtained from ATCC and maintained in high glucose DMEM supplemented with GlutaMAX (Thermo Fisher Scientific, 61965059) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco, 26140079), 1 mM sodium pyruvate, 100 U/ml penicillin and 100 μg/ml streptomycin (Thermo Fisher Scientific, 15140-122) and cultured at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Immunostaining, Derivative Assay, Transfection, Control, Fluorescence, Two Tailed Test

Journal: Journal of Cell Science

Article Title: Selective disruption of microtubule formation at the nuclear envelope impairs the bone resorption capacity of osteoclasts

doi: 10.1242/jcs.264166

Figure Lengend Snippet: AKAP6 depletion disrupts microtubule-actin cytoskeletal crosstalk in osteoclasts. (A) Representative images of RAW264.7-derived osteoclasts transfected with siControl or siAKAP6, stained for F-actin (phalloidin, green) and microtubules (α-tubulin, red). Corresponding BIOP JACoP analyses are shown in grayscale, illustrating the spatial overlap between F-actin and microtubules (lower panel). Scale bars: 10 μm. Dotted white lines indicate the boundary between the actin-dense sealing zone and the cytoplasm. (B) Quantification of microtubule-actin overlap in siControl and siAKAP6-treated osteoclasts. The actin-microtubule overlap was quantified in arbitrary units (a.u.) as the ratio of actin that colocalized with microtubules normalized against the total actin pixel count. SuperPlots display the overall mean (black line) and biological-replicate mean (color-coded triangles) from n =3, with the individual cells (12–25 per condition) shown as matching color-coded dots superimposed beneath them. ** P =0.0043 (paired two-tailed t -test). (C) Representative immunostaining images of RAW264.7-derived osteoclasts transfected with siControl or siAKAP6 co-stained for actin (phalloidin, green) and microtubules (α-tubulin, red). The right panels show higher magnification images of the boxed regions, highlighting actin ring structure. Images in C representative of three experimental repeats. Scale bars: 10 μm.

Article Snippet: RAW264.7 cells were obtained from ATCC and maintained in high glucose DMEM supplemented with GlutaMAX (Thermo Fisher Scientific, 61965059) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco, 26140079), 1 mM sodium pyruvate, 100 U/ml penicillin and 100 μg/ml streptomycin (Thermo Fisher Scientific, 15140-122) and cultured at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Derivative Assay, Transfection, Staining, Two Tailed Test, Immunostaining

Journal: Journal of Cell Science

Article Title: Selective disruption of microtubule formation at the nuclear envelope impairs the bone resorption capacity of osteoclasts

doi: 10.1242/jcs.264166

Figure Lengend Snippet: AKAP6 depletion impairs osteoclast resorptive activity in RAW264.7-derived osteoclasts. (A) Representative images of RAW264.7-derived osteoclasts transfected with siControl- and siAKAP6 cultured on CaP-coated wells. Cells were removed, and the CaP substrate was stained with 2.5% AgNO 3 to visualize calcium deposits; resorption pits appear as unstained areas. Scale bars: 250 µm. (B) Relative resorbed area of siControl and siAKAP6-treated osteoclasts. Data represent mean±s.d. of three independent experiments, with three wells per experiment. **P =0.0081 (paired two-tailed t -test). (C) Representative images of RAW264.7-derived osteoclasts transfected with siControl or siAKAP6, and stained for F-actin (phalloidin, red), CaP (calcein, green) and nuclei (DAPI, blue). Note, calcein stains CaP, meaning resorption pits with lower calcein intensity indicate higher osteoclast resorption. Scale bars: 10 μm. (D) Quantification of osteoclast resorptive activity across sealing zone size categories (<500 μm 2 , 500–2000 μm 2 , >2000–5000 μm 2 and >5000 μm 2 ). SuperPlots display individual resorption pits as color-coded dots (siControl, blue; siAKAP6, red), with biological replicate means ( n =3; 35–80 pits per experiment, equal numbers per condition) superimposed as colored triangles, each color representing a different replicate. Error bars show overall mean±s.d. Resorption was quantified using the calcein ratio (calcein intensity inside the sealing zone divided by the intensity outside), where values <1 indicate active resorption. *P <0.05; **P <0.01 (two-way ANOVA with Bonferroni's post-hoc test).

Article Snippet: RAW264.7 cells were obtained from ATCC and maintained in high glucose DMEM supplemented with GlutaMAX (Thermo Fisher Scientific, 61965059) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco, 26140079), 1 mM sodium pyruvate, 100 U/ml penicillin and 100 μg/ml streptomycin (Thermo Fisher Scientific, 15140-122) and cultured at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Activity Assay, Derivative Assay, Transfection, Cell Culture, Staining, Two Tailed Test

Journal: Bioactive Materials

Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis

doi: 10.1016/j.bioactmat.2026.01.001

Figure Lengend Snippet: (A–C) Cell adhesion, relative fluorescence intensity of CCR7/CD206 and ROS for Raw264.7 on different Gaussian curvatures. (D) CCR7 & CD206 staining of Raw264.7 cells (blue-nucleus, red-CCR7, green-CD206, scale:100 μm). (E, F) F-actin & DAPI staining and statistical scatterplot of hBMSC adhesion. (G) ALP intensity of hBMSCs on different Gaussian curvatures. (H, I) Cell adhesion and CD31 fluorescence intensity of HUVECs. (J) Chord diagram for comprehensive normalized data of Raw264.7, hBMSCs and HUVECs behavior detection with different Gaussian curvatures on HCGC chips.

Article Snippet: The cells types utilized in this study included: hBMSCs (No. 7500, ScienCell, USA.), Raw264.7 (CL-0190, Procell system, China), HUVECs (No. 8000, ScienCell, USA.), rBMSCs (MUBMX-01001, OriCell, USA.) and MS1(CL-0162, Procell system, China).

Techniques: Fluorescence, Staining

Journal: Bioactive Materials

Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis

doi: 10.1016/j.bioactmat.2026.01.001

Figure Lengend Snippet: Macrophage polarization analysis of Raw264.7 on structures with different gaussian curvature: (A, B) Chord Diagram for qPCR analysis of CCR7, IL6, iNOS-inflammatory and M1 marker genes, and Arg-1, CD206, IL10-M2 related protein genes in different Gaussian curvature groups. (C) Protein content of Arg-1 in different Gaussian curvature groups at 1 and 3 days. (D) Integral plots of the five experimental groups. IL4 group is the positive control for CD206 expression and lipopolysaccharide (LPS) group is the negative control.

Article Snippet: The cells types utilized in this study included: hBMSCs (No. 7500, ScienCell, USA.), Raw264.7 (CL-0190, Procell system, China), HUVECs (No. 8000, ScienCell, USA.), rBMSCs (MUBMX-01001, OriCell, USA.) and MS1(CL-0162, Procell system, China).

Techniques: Marker, Positive Control, Expressing, Negative Control

Journal: Bioactive Materials

Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis

doi: 10.1016/j.bioactmat.2026.01.001

Figure Lengend Snippet: RNA-seq analysis of relevant genes in Raw264.7 cells on various Gaussian curvature: (A) Top 20 functional molecule enrichment of differential genes. (B) KEGG signaling pathway enrichment analysis. (C–G) The gene expression heat map of macrophage polarization, HIF-1α, Ras-MAPK signaling pathway, Focal adhesion and microtubule binding. (H) Potential mechanism of HIF-1α Downregulation by Negative Gaussian Curvature via Ras-MAPK Pathway.

Article Snippet: The cells types utilized in this study included: hBMSCs (No. 7500, ScienCell, USA.), Raw264.7 (CL-0190, Procell system, China), HUVECs (No. 8000, ScienCell, USA.), rBMSCs (MUBMX-01001, OriCell, USA.) and MS1(CL-0162, Procell system, China).

Techniques: RNA Sequencing, Functional Assay, Gene Expression, Binding Assay

Journal: Bioactive Materials

Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis

doi: 10.1016/j.bioactmat.2026.01.001

Figure Lengend Snippet: (A) Schematic diagram of transwell migration of rBMSCs and MS1 co-cultured with Raw264.7. (B) BMP-2 concentration in Raw264.7-CM. (C)The statistics of rBMSCs migration through transwell. (D, E) ALP staining for 7 days and ALP quantification of rBMSCs incubated with Raw264.7-CM for 7 days/14 days. (F) Gene Expression of Col-I, RunX2, and ALP in rBMSCs incubated with Raw264.7-CM. (G) VEGF concentration in Raw264.7-CM. (H) Transwell migration bar statistics (n = 3, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001). (I) Gene expression of VEGF and TGF-β in MS1 incubated with Raw264.7-CM. (J) Fluorescence image of MS1 incubated with Raw264.7-CM in tube formation assay , the Raw264.7-CM collected from Raw264.7 on the plate was the blank.

Article Snippet: The cells types utilized in this study included: hBMSCs (No. 7500, ScienCell, USA.), Raw264.7 (CL-0190, Procell system, China), HUVECs (No. 8000, ScienCell, USA.), rBMSCs (MUBMX-01001, OriCell, USA.) and MS1(CL-0162, Procell system, China).

Techniques: Migration, Cell Culture, Concentration Assay, Staining, Incubation, Gene Expression, Fluorescence, Tube Formation Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Elevated IgG levels drive macrophage M2-to-M1 reprogramming. (A) Sphenoid sinus-invasive tumor cases stratified into CD19-high (n=5) and CD19-low (n=5) groups based on the cohort median of CD19 + B cell density, with (B) quantitative analyses of macrophage polarization (M1-like versus M2-like). (C) Dural-invasive tumor and non-invasive tumor cases stratified into IgG-high (n=27) and IgG-low (n=26) groups based on the cohort median of relative IgG immunohistochemistry staining intensity, with (D) quantitative analyses of M1-like/M2-like macrophage proportions. (E and F) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by IgG (10 µg/ml) exposure. Relative (E) IL-6 and (F) TNF-α mRNA expression in RAW264.7 macrophages pre-polarized to M0, M1 or M2 states. (G) Representative flow cytometric cell-cycle profiles of TtT/GF cells following the indicated treatments. (H) Stacked bar plot summarizing the percentages of cells from (G) in G 1 , S and G 2 /M phases. (I) Representative images from the scratch wound assay at 0, 24, 48 and 72 h under the indicated treatments. (J) Quantification of scratch wound closure. (K) Representative western blot images showing total STAT1, p-STAT1, total STAT3, p-STAT3 and β-actin levels in cells treated with IFN-γ (100 ng/ml), IL-6 (100 ng/ml), IFN-γ + IL-6 (50 ng/ml each), ruxolitinib (5 µM) or IFN-γ + IL-6 (50 ng/ml each) plus ruxolitinib (5 µM), as indicated. (L) Densitometric semi-quantification of p-STAT1/STAT1 (ratio). (B and D) Unpaired two-tailed Student's t-test. (E, F, J and L) One-way ANOVA with Tukey's post hoc multiple comparisons test. *P<0.05, ***P<0.001, ****P<0.0001. CTRL, control; IBA-1, ionised calcium binding adaptor molecule 1; ns, not significant; p-, phosphorylated; PE-A, phycoerythrin-area.

Article Snippet: RAW264.7 mouse macrophages (ATCC ® TIB-71TM; American Type Culture Collection) were maintained in complete DMEM at 37°C with 5% CO 2 .

Techniques: Immunohistochemistry, Staining, Expressing, Scratch Wound Assay Assay, Western Blot, Two Tailed Test, Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Anti-CD47 mAb enhances ADCP to suppress tumor cell proliferation. (A) Immunofluorescence staining of CD47 (red) and DAPI (blue) in a representative subset of non-invasive tumor, dural-invasive tumor and sphenoid sinus-invasive tumor cases (n=10 per group). (B) Paired comparison of CD47 fluorescence intensity at the IF versus the TC. (C) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by anti-CD47 mAb (10 µg/ml) treatment for 12 h. Quantitative PCR was used to analyze polarization/activation markers. (D) Schematic illustrating anti-CD47 mAb-mediated blockade of the CD47-SIRPα axis and enhancement of ADCP. (E) EdU assay of TtT/GF cell proliferation in a Transwell co-culture with anti-CD47 mAb-treated polarized macrophages. (F) Quantification of EdU-positive cells. (G) Representative microscopy images and flow cytometry plots showing macrophage phagocytosis of pHrodo™ Red-labeled GFP-TtT/GF cells. (H) Quantification of phagocytosis (%). (B) Paired two-tailed Student's t-test. (C, F and H) One-way ANOVA with Tukey's post hoc multiple comparisons test. **P<0.01, ***P<0.001, ****P<0.0001. ADCP, antibody-dependent cellular phagocytosis; Arg-1, arginase 1; EdU, 5-ethynyl-2′-deoxyuridine; FcγR, Fcγ receptor; GFP, green fluorescent protein; IF, invasive front; mAb, monoclonal antibody; NOS2, nitric oxide synthase 2; ns, not significant; PE, phycoerythrin; SIRPα, signal regulatory protein-α; SSCA, side scatter area; TC, tumor core.

Article Snippet: RAW264.7 mouse macrophages (ATCC ® TIB-71TM; American Type Culture Collection) were maintained in complete DMEM at 37°C with 5% CO 2 .

Techniques: Immunofluorescence, Staining, Comparison, Fluorescence, Real-time Polymerase Chain Reaction, Activation Assay, EdU Assay, Co-Culture Assay, Microscopy, Flow Cytometry, Labeling, Two Tailed Test

Journal: Bioactive Materials

Article Title: Integrated fabrication of a shape-adaptable, antioxidative composite stent for effective closure and biological repair of enteroatmospheric fistula

doi: 10.1016/j.bioactmat.2026.01.014

Figure Lengend Snippet: Antioxidant and anti-inflammatory effects of the composite stent. (A) ROS levels in RAW264.7 and IEC-6 cells by DCFH-DA staining. (B, C) Quantification of ROS fluorescence by integrated density. (D) Mitochondrial ROS detected by MitoSOX staining. (E, F) Quantification of mitochondrial ROS by integrated density. (G) Co-culture system of the composite stent with macrophages using Transwell chambers. (H) Flow cytometry analysis of macrophage polarization based on CD86 (M1 marker) and CD206 (M2 marker) expression under different stimuli. (I) Quantitative analysis of CD86 + macrophages obtained from flow cytometry. (J–L) ELISA measurements of pro-inflammatory cytokines (J) TNF-α, (K) IL-6, and (L) IFN-β in the culture supernatant. Data were presented as mean ± SD (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: IEC-6 cells and RAW264.7 cells were purchased from KeyGEN BioTech (Nanjing, China).

Techniques: Staining, Fluorescence, Co-Culture Assay, Flow Cytometry, Marker, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Materials Today Bio

Article Title: Attenuating the pathological cycle of periodontitis: CGL NPs suppress pathological mineralization and modulate the periodontal microenvironment

doi: 10.1016/j.mtbio.2026.103156

Figure Lengend Snippet: Biocompatibility of CGL NPs. (A, B) Cell viability of HGFs (A) and RAW264.7 cells (B) assessed by CCK-8 assay. (C, D) Live/dead fluorescence staining of HGFs (C) and RAW264.7 cells (D). (E, F) Transwell migration assay images (E) and quantitative analysis (F) of HGFs. (G, H) Wound healing assay images (G) and quantitative analysis (H) of HGFs. (I) LDH release assay quantifying cytotoxicity in RAW264.7 cells. (∗ denotes that the difference exhibits statistical significance when compared with the blank group. Data are presented as mean ± standard deviation for n = 3. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, ns denotes the absence of a statistically significant difference.)

Article Snippet: RAW264.7 cells and HGFs were obtained from Servicebio Biotechnology Co., Ltd. (Wuhan, China) and cultured in DMEM (Gibco, USA) supplemented with 10% FBS (Gibco, USA) and 1% penicillin-streptomycin (Beyotime, China) in a humidified incubator at 37 °C with 5% CO2.

Techniques: CCK-8 Assay, Fluorescence, Staining, Transwell Migration Assay, Wound Healing Assay, Lactate Dehydrogenase Assay, Standard Deviation