Journal: BMC Pulmonary Medicine

Article Title: Optimization of intranasal bleomycin dose for effective pulmonary fibrosis induction in mice with minimal animal distress

doi: 10.1186/s12890-025-04001-4

Figure Lengend Snippet: Histopathological characterization in BLM-treated animals (IN—5 mg/kg). A ) Histopathological evaluation by H&E staining (upper panel), Sirius Red staining (middle panel), and Masson’s trichrome (lower panel) of lungs of vehicle mice (CTR) or treated with BLM after a single IN administration. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported. Scale bar = 500 µm (upper panel) – 100 µm (lower panels). B ) IHC for Iba1 (left panel), CD68 (green—middle panel), and α-SMA staining (right panel) of lungs of CTR or treated with BLM. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported, scale bar = 100 µm. The boxed areas (Iba1 black, CD68 red, α-SMA green) in CTR and day 7 sections are shown at higher magnification in the right panel, scale bar = 50 µm. C) Histopathological quantification of Sirius Red staining in lung section (left graph) and representation of Ashcroft scale grade (right graph) obtained by Masson’s trichrome analysis of lungs of CTR or treated with BLM. Data are reported as mean ± SE. The data were analyzed by Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, ** p ≤ 0.01. D ) mRNA expression of TNF-α, COL 1a1, and FN1 was evaluated by RT-qPCR in the lungs of mice ( n = 3 per group) treated with BLM and sacrificed at different time points. Genes were normalized on β-ACT, and the 2 −ΔΔCt method was employed for relative quantification on an external calibrator. Data are reported as mean ± SE and were analyzed with Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, * p ≤ 0.05, ** p ≤ 0.01. E ) SMAD 2/3 and pSMAD 2 expression in the lungs from CTR mouse at days 7, 14, 21, and 28 of treatment obtained with WB

Article Snippet: For subcellular localization, primary rat anti-CD68 monoclonal antibody (1:200, Serotec, Kidlington, UK) + Triton X-100 0.1% + NGS 3% in 1X PBS overnight at 4 °C was used.

Techniques: Staining, Expressing, Quantitative RT-PCR, Quantitative Proteomics

Journal: BMC Pulmonary Medicine

Article Title: Optimization of intranasal bleomycin dose for effective pulmonary fibrosis induction in mice with minimal animal distress

doi: 10.1186/s12890-025-04001-4

Figure Lengend Snippet: Histopathological characterization in BLM-treated animals (IN—3 mg/kg). A Histopathological evaluation by H&E staining (upper panel), Sirius Red staining (middle panel), and Masson’s trichrome (lower panel) of lungs of vehicle mice (CTR) or treated with BLM after a single IN administration. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported. Scale bar = 500 µm (upper panel) – 100 µm (lower panels). B IHC for Iba1 (left panel), CD68 (green—middle panel), and α-SMA staining (right panel) of lungs of CTR or treated with BLM. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported. Scale bar = 100 µm. The boxed areas (Iba1 black, CD68 red, α-SMA green) in CTR and day 7 sections are shown at higher magnification in the right panel, scale bar = 50 µm C ) Histopathological quantification of Sirius Red staining in lung section (left graph) and representation of Ashcroft scale grade (right graph) obtained by Masson’s trichrome analysis of lungs of CTR or treated with BLM. Data are reported as mean ± SE. The data were analyzed by Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. D ) mRNA expression of TNF-α, COL 1a1, and FN1 was evaluated by RT-qPCR in the lungs of mice ( n = 3 per group) treated with BLM and sacrificed at different time points. Genes were normalized on β-actin, and the 2 −ΔΔCt method was employed for relative quantification on an external calibrator. Data are reported as mean ± SE and were analyzed with Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, * p ≤ 0.05, ** p ≤ 0.01. E) SMAD 2/3 and pSMAD 2 expression in the lungs from CTR mouse at days 7, 14, 21, and 28 of treatment obtained with WB

Article Snippet: For subcellular localization, primary rat anti-CD68 monoclonal antibody (1:200, Serotec, Kidlington, UK) + Triton X-100 0.1% + NGS 3% in 1X PBS overnight at 4 °C was used.

Techniques: Staining, Expressing, Quantitative RT-PCR, Quantitative Proteomics

Journal: bioRxiv

Article Title: Targeting Macrophage Folate Receptor-β for Diagnosis and Treatment of Autoimmune Myocarditis

doi: 10.1101/2025.11.02.686077

Figure Lengend Snippet: ( A ) Representative images of hematoxylin-eosin (H&E) staining and immunohistochemistry for CD68-positive macrophages in rat hearts collected 21 days after the first immunization. Rats were administered EC2319 or saline throughout the induction of myocarditis. ( B ) Quantification of CD68-positive macrophages in myocardial tissue on Day 21 of the prevention sub-study.

Article Snippet: For immunohistochemistry, adjacent serial 8-μm cryosections were stained with monoclonal mouse anti-rat CD68 antibody (1:1000, MCA341GA, Bio-Rad) to identify macrophages.

Techniques: Staining, Immunohistochemistry, Saline

Journal: bioRxiv

Article Title: Targeting Macrophage Folate Receptor-β for Diagnosis and Treatment of Autoimmune Myocarditis

doi: 10.1101/2025.11.02.686077

Figure Lengend Snippet: ( A ) Representative [ 18 F]FOL autoradiographs and corresponding hematoxylin and eosin (H&E) and immunohistochemical stainings of myocarditis rat hearts post-treatment. ( B ) Quantification of CD68-positive macrophages and ( C ) correlation with myocardial [ 18 F]FOL uptake on Day 26. Spearman correlation coefficient was used to analyze linear correlations between CD68-positive area and myocardial [ 18 F]FOL uptake on Day 26. ( D ) Quantifications of M1-polarized macrophages (iNOS-positive), ( E ) M2-polarized macrophages (MRC-1-positive), and ( F ) M1/M2 macrophage ratios in inflammatory lesions based on immunostaining. ( G ) Quantification of fibrotic area using Masson’s trichrome staining. n = 6-8 per group for B-C; n = 3 per group for D-F; n = 5-7 per group for G. Statistical comparisons of normally distributed data were performed using one-way ANOVA followed by post hoc tests: Tukey’s test (for homogeneous variances) and Games–Howell test (for non-homogeneous variances) to compare different treatments. Non-normally distributed continuous data are presented as median (interquartile range, IQR) and compared using the Kruskal–Wallis test, followed by pairwise comparisons with Bonferroni-adjusted p -values.

Article Snippet: For immunohistochemistry, adjacent serial 8-μm cryosections were stained with monoclonal mouse anti-rat CD68 antibody (1:1000, MCA341GA, Bio-Rad) to identify macrophages.

Techniques: Immunohistochemical staining, Immunostaining, Staining

Journal: bioRxiv

Article Title: Targeting Macrophage Folate Receptor-β for Diagnosis and Treatment of Autoimmune Myocarditis

doi: 10.1101/2025.11.02.686077

Figure Lengend Snippet: There was strong co-localization of CD68 and iNOS, a marker of pro-inflammatory M1 macrophages, whereas there was only partial co-localization of CD68 and MRC-1, a marker of anti-inflammatory M2 macrophages. Number of iNOS-positive macrophages is markedly lower after treatment with EC2319 than saline or Etanercept, consistent with a shift toward an anti-inflammatory macrophage profile.

Article Snippet: For immunohistochemistry, adjacent serial 8-μm cryosections were stained with monoclonal mouse anti-rat CD68 antibody (1:1000, MCA341GA, Bio-Rad) to identify macrophages.

Techniques: Marker, Saline

Journal: Acta Neuropathologica Communications

Article Title: Blocking axon-glial mechanotransduction to prevent concussive brain injury

doi: 10.1186/s40478-025-02117-6

Figure Lengend Snippet: Memantine inhibits CHIMERA-induced microglial activation and cortical demyelination but not axonal varicosity formation. ( A ) CHIMERA-induced microglial activation in the cortex. Low-mag images of the cortex from mice received no impact (Sham, left), or 0 h, 4 h, 24 h, or 3d after head impact. The anti-CD68 staining signals are in red, YFP in green, and Hoechst in blue. ( B ) Summary of the density of CD68 + cells in the mouse cortex under different conditions. Mouse numbers are provided within the bars. 3–4 images from each mouse are included. One-way ANOVA followed by Dunnett’s test: * p < 0.05, *** p < 0.001. ( C ) Diagram for the memantine experiment. Memantine was injected (via i.p. at 10 mg/kg each dose) 1.5 h before CHIMERA and 3 h afterward. ( D ) Representative images of induced axonal varicosities at 0 h from vehicle (top) and memantine (bottom) treated Thy1-YFP transgenic mice. YFP fluorescence signals are in green, the CD68 signals in red, and Hoechst in blue. ( E ) Summary of the effect of memantine treatment on axonal varicosity induction in the cortex (left) and EC and CC (right) of Thy1-YFP transgenic mouse at 0 h and 24 h. Green lines, the basal levels in Sham. Unpaired t-test: no significant difference. n = 6 for all conditions. ( F ) Memantine treatment eliminated CD68 + cells in the cortex and significantly reduced CD68 staining signals in the EC 24 h after CHIMERA. CD68 staining signals are inverted in grayscale images. High-mag confocal images are on the right. ( G ) Memantine treatment markedly reduced CD68 + cells in the CC 24 h after CHIMERA. High-mag confocal images are at the bottom. ( H ) Summary of memantine’s effect on the density of CHIMERA-induced CD68 + cells at 24 h. Unpaired t-test: ** p < 0.01, *** p < 0.001. ( I ) Memantine treatment markedly reduced dMBP signals in the cortex 24 h after CHIMERA. Unpaired t-test: *** p < 0.001. n = 6 for all conditions. ( J ) Confocal image stacks of CD68 (green) and dMBP (red) costaining in the cortex of vehicle (left) and memantine-treated mice 24 h after CHIMERA. Scale bars, 250 μm in ( A ), ( F ) left, ( G ) upper, and ( I ); 25 μm in ( D ), ( F ) right, ( G ) lower, and ( J )

Article Snippet: Antibodies used in this study include a rabbit polyclonal anti-degraded MBP antibody that recognizes damaged MBP proteins (1:2000, anti-dMBP; Catalog #: AB5864; Chemicon), a rat monoclonal anti-MBP antibody (1:200; Catalog #: ab7349; abcam), a rabbit polyclonal anti-TRPV4 antibody (1:200; Catalog #: LS-A8583; LifeSpan BioSciences, Seattle, Washington, USA), rabbit polyclonal anti-Cav2.1, anti-Cav2.2 and anti-Cav2.3 antibodies (Catalog #s: ACC-001, ACC-002 and ACC-006; Alomone Labs, Jerusalem, Israel), a goat polyclonal anti-GFAP antibody (1:200; Catolog #: Ab53554; Abcam Inc. Waltham, MA, USA), and a rat monoclonal anti-CD68 antibody (1:200; Catalog #: MCA1957GA; Bio-Rad; Hercules, CA, USA), as well as Cy3 and Cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Techniques: Activation Assay, Staining, Injection, Transgenic Assay, Fluorescence

Journal: Acta Neuropathologica Communications

Article Title: Blocking axon-glial mechanotransduction to prevent concussive brain injury

doi: 10.1186/s40478-025-02117-6

Figure Lengend Snippet: GSK219 markedly inhibits CHIMERA-induced axonal varicosities, microglial activation, cortical demyelination, and behavioral alterations. ( A ) Diagram for the CHIMERA experiment with GSK219 pretreatment. Mice were gavaged with GSK219 (20 mg/kg) or vehicle (as control) once 3 h before CHIMERA (0.9 J). ( B )-( C ) Confocal images of the cortex of Thy1-YFP mice 24 h after CHIMERA with the pretreatment of vehicle ( B ) or GSK219 ( C ). The dMBP staining signals are inverted in grayscale images (right) and in red in merged images (left), YFP in green, and Hoechst in blue. ( D ) Summary of YFP + axons with varicosities in EC 0 h after CHIMERA with vehicle or GSK219 pretreatment. The green line, the basal level in Sham. The mouse numbers are provided in the chart. 1–3 images from each mouse are included. Unpaired t-test: p = 0.0000021. ( E ) Summary of dMBP intensity in the cortex 24 h after CHIMERA with vehicle or GSK219 pretreatment. The green line, the basal level in Sham. Unpaired t-test: p = 7.481 × 10 − 12 . ( F ) Summary of CD68 + cell density in the cortex 24 h after CHIMERA with vehicle or GSP219 pretreatment. The basal level in Sham is close to 0. Unpaired t-test: p = 3.257 × 10 − 14 . ( G ) Example traces of mouse movement in the elevated plus maze (EPM) before (cyan) or 3d after CHIMERA (black) with vehicle (left) or GSK219 pretreatment (right). ( H ) Summary of the percentage time of open arms, 100 × T open /(T open +T closed ), in the EPM test. Unpaired t-test: p = 0.00408 at 3 DPI. ( I ) Summary of the total travel distance in the EPM test. Unpaired t-test: p = 0.0498 at 14 DPI. ( J ) Rotarod results before CHIMERA (-3d), and 3d, 7d, 14d, and 30d after CHIMERA. Unpaired t-test in 3d: trial 2 p = 0.0157; trial 3 p = 0.00606; trial 4 p = 0.000829; trial 5 p = 0.00133. ( K ) Diagram for the CHIMERA experiment with GSK219 post-treatment. Mice were gavaged with GSK219 (20 mg/kg) or vehicle (as control) once 2 h after CHIMERA (0.9 J). ( L ) Summary of YFP + axons with varicosities in EC 24 h after CHIMERA with vehicle or GSK219 post-treatment. The green line, the basal level in Sham. Unpaired t-test: p = 0.000205. ( M ) Summary of dMBP intensity in the cortex 24 h after CHIMERA with vehicle or GSK219 post-treatment. The green line, the basal level in Sham. Unpaired t-test: p = 0.0000517. ( N ) Summary of CD68 + cell density in the cortex 24 h after CHIMERA with vehicle or GSK219 post-treatment. The basal level in Sham is close to 0. p = 0.957. Unpaired t-test: * p < 0.05, ** p < 0.01, *** p < 0.001. Scale bars, 20 μm in ( B ) and ( C )

Article Snippet: Antibodies used in this study include a rabbit polyclonal anti-degraded MBP antibody that recognizes damaged MBP proteins (1:2000, anti-dMBP; Catalog #: AB5864; Chemicon), a rat monoclonal anti-MBP antibody (1:200; Catalog #: ab7349; abcam), a rabbit polyclonal anti-TRPV4 antibody (1:200; Catalog #: LS-A8583; LifeSpan BioSciences, Seattle, Washington, USA), rabbit polyclonal anti-Cav2.1, anti-Cav2.2 and anti-Cav2.3 antibodies (Catalog #s: ACC-001, ACC-002 and ACC-006; Alomone Labs, Jerusalem, Israel), a goat polyclonal anti-GFAP antibody (1:200; Catolog #: Ab53554; Abcam Inc. Waltham, MA, USA), and a rat monoclonal anti-CD68 antibody (1:200; Catalog #: MCA1957GA; Bio-Rad; Hercules, CA, USA), as well as Cy3 and Cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Techniques: Activation Assay, Control, Staining

Journal: Acta Neuropathologica Communications

Article Title: Blocking axon-glial mechanotransduction to prevent concussive brain injury

doi: 10.1186/s40478-025-02117-6

Figure Lengend Snippet: Acute deletion of neuronal TRPV4 prevents CHIMERA-induced axonal varicosity formation and adjacent glial changes. ( A ) Diagram of injecting AAV9-hSyn-Cre-dTomato into the right cortex of mice. Adult WT and TRPV4 fl/fl mice were used in CHIMERA about 1 month after the viral injection. A low-mag image (bottom) shows the injection sites with dTomato fluorescence in green and Hoechst in blue. ( B ) Low-mag gray-scale images with dTomato signals inverted of the cortex and EC from a WT (top) or a TRPV4 fl/fl (bottom) mouse injected with AAV9:Syn-Cre; dTomato at 0 h after CHIMERA. ( C ) Confocal images from the EC of the mice in ( B ). The dTomato signals are in green and Hoechst in blue. ( D ) Summary of the percentage of dTomato + axons with varicosities in white matter (CC and EC) and gray matter (cortical layers 1–6) from WT (black) and TRPV4 fl/fl (red) mice 0–24 h after CHIMERA, compared to Sham. n = 7 for all conditions. ( E ) Confocal images and quantification of TRPV4 protein (red) knockdown via the infection by AAV9-Syn-Cre-dTomato (green) in the cortex of TRPV4 fl/fl but not WT mice. Unpaired t-test: **, p < 0.01. n = 7 for both conditions. ( F ) Confocal images from the Ipsilateral and Contralateral sides of the cortex from WT (top) or TRPV4 fl/fl (bottom) mice 24 h after CHIMERA. The dTomato signals are in green and dMBP signals are in red. ( G ) Summary of dMBP staining intensity from the mouse cortex under different conditions. ( H ) Summary of CD68 + cell density in the cortex from WT or TRPV4 fl/fl (Flox) mice with AAV injection 24 h after CHIMERA. n = 10 in all groups. Unpaired t-test: **, p < 0.01; ***, p < 0.001. Scale bars, 160 μm in ( B ); 40 μm in ( C ), ( E ), and ( F )

Article Snippet: Antibodies used in this study include a rabbit polyclonal anti-degraded MBP antibody that recognizes damaged MBP proteins (1:2000, anti-dMBP; Catalog #: AB5864; Chemicon), a rat monoclonal anti-MBP antibody (1:200; Catalog #: ab7349; abcam), a rabbit polyclonal anti-TRPV4 antibody (1:200; Catalog #: LS-A8583; LifeSpan BioSciences, Seattle, Washington, USA), rabbit polyclonal anti-Cav2.1, anti-Cav2.2 and anti-Cav2.3 antibodies (Catalog #s: ACC-001, ACC-002 and ACC-006; Alomone Labs, Jerusalem, Israel), a goat polyclonal anti-GFAP antibody (1:200; Catolog #: Ab53554; Abcam Inc. Waltham, MA, USA), and a rat monoclonal anti-CD68 antibody (1:200; Catalog #: MCA1957GA; Bio-Rad; Hercules, CA, USA), as well as Cy3 and Cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Techniques: Injection, Fluorescence, Knockdown, Infection, Staining

Journal: Acta Neuropathologica Communications

Article Title: Blocking axon-glial mechanotransduction to prevent concussive brain injury

doi: 10.1186/s40478-025-02117-6

Figure Lengend Snippet: GSK279 markedly inhibits CHIMERA-induced axon-glial changes in WT but not TRPV4 −/− mice. ( A ) Structural diagrams of GSK219 and GSK279. ( B ) Diagram for the CHIMERA experiment with GSK279 pretreatment. WT; Thy1-YFP or TRPV4 −/− ;Thy1-YFP mice (3–4 months old) were injected via tail vein with GSK279 (18 µg/kg) or vehicle (as control) once 1.5 h before CHIMERA (0.9 J). Mice were perfused and fixed either immediately (0 h) or 24 h after head impact. ( C ) Summary of YFP + axons with varicosities in EC 0 h after CHIMERA with vehicle or GSK279 pretreatment. The green line, the basal level in Sham. ( D ) Confocal images of the EC of WT (left) and TRPV4 −/− (right) mice 24 h after CHIMERA with the pretreatment of vehicle (top) or GSK279 (bottom). The CD68 staining signals are inverted in grayscale images (right) and red in merged images (left), YFP in green, and Hoechst in blue. ( E ) Summary of YFP + axons with varicosities in EC 24 h after CHIMERA. ( F ) Summary of CD68 + cell density in the cortex 24 h after CHIMERA. ( G ) The dMBP staining signals (inverted in gray-scale images (right) and in red in merged images (left) in the cortex of WT mice 24 h after CHIMERA with vehicle (top) or GSP279 (middle) pretreatment. The summary is at the bottom. ( H ) The dMBP staining signals in the cortex of TRPV4 −/− mice 24 h after CHIMERA with vehicle or GSK279 pretreatment. Mouse numbers, n = 3 for all groups. 1–3 images from each mouse are included. Unpaired t-test: ***, p < 0.001. Scale bars, 20 μm in ( D ), ( G ), and ( H )

Article Snippet: Antibodies used in this study include a rabbit polyclonal anti-degraded MBP antibody that recognizes damaged MBP proteins (1:2000, anti-dMBP; Catalog #: AB5864; Chemicon), a rat monoclonal anti-MBP antibody (1:200; Catalog #: ab7349; abcam), a rabbit polyclonal anti-TRPV4 antibody (1:200; Catalog #: LS-A8583; LifeSpan BioSciences, Seattle, Washington, USA), rabbit polyclonal anti-Cav2.1, anti-Cav2.2 and anti-Cav2.3 antibodies (Catalog #s: ACC-001, ACC-002 and ACC-006; Alomone Labs, Jerusalem, Israel), a goat polyclonal anti-GFAP antibody (1:200; Catolog #: Ab53554; Abcam Inc. Waltham, MA, USA), and a rat monoclonal anti-CD68 antibody (1:200; Catalog #: MCA1957GA; Bio-Rad; Hercules, CA, USA), as well as Cy3 and Cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Techniques: Injection, Control, Staining

Journal: Acta Neuropathologica Communications

Article Title: Blocking axon-glial mechanotransduction to prevent concussive brain injury

doi: 10.1186/s40478-025-02117-6

Figure Lengend Snippet: Gabapentin markedly inhibits CHIMERA-induced axon-glial changes in TRPV4 −/− mice. ( A ) Diagram for the CHIMERA experiment with gabapentin pretreatment. WT; Thy1-YFP or TRPV4 −/− ;Thy1-YFP mice were injected via I.P. with gabapentin (50 mg/kg) or vehicle (as control) once 1.5 h before CHIMERA (0.9 J). Mice were perfused and fixed either immediately (0 h) or 24 h after head impact. ( B ) Summary of YFP + axons with varicosities in EC 0 h after CHIMERA with vehicle or gabapentin pretreatment. The green line, the basal level in Sham. ( C ) Summary of YFP + axons with varicosities in EC 24 h after CHIMERA. ( D ) Confocal images of the EC of WT (left) and TRPV4 −/− (right) mice 24 h after CHIMERA with the pretreatment of vehicle (Veh; top) or gabapentin (Gab; bottom). The CD68 staining signals are inverted in grayscale images (right) and in red in merged images (left), YFP in green, and Hoechst in blue. ( E ) Summary of CD68 + cell density in the cortex 24 h after CHIMERA. ( F ) The dMBP staining signals in the cortex of WT (left) or TRPV4 −/− (right) mice 24 h after CHIMERA. ( G ) Summary of dMBP signals in the cortex 24 h after CHIMERA. Mouse numbers, n = 3 for all groups. 1–3 images from each mouse are included. Unpaired t-test: **, p < 0.01; ***, p < 0.001. Scale bars, 25 μm in ( D ) and ( F )

Article Snippet: Antibodies used in this study include a rabbit polyclonal anti-degraded MBP antibody that recognizes damaged MBP proteins (1:2000, anti-dMBP; Catalog #: AB5864; Chemicon), a rat monoclonal anti-MBP antibody (1:200; Catalog #: ab7349; abcam), a rabbit polyclonal anti-TRPV4 antibody (1:200; Catalog #: LS-A8583; LifeSpan BioSciences, Seattle, Washington, USA), rabbit polyclonal anti-Cav2.1, anti-Cav2.2 and anti-Cav2.3 antibodies (Catalog #s: ACC-001, ACC-002 and ACC-006; Alomone Labs, Jerusalem, Israel), a goat polyclonal anti-GFAP antibody (1:200; Catolog #: Ab53554; Abcam Inc. Waltham, MA, USA), and a rat monoclonal anti-CD68 antibody (1:200; Catalog #: MCA1957GA; Bio-Rad; Hercules, CA, USA), as well as Cy3 and Cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Techniques: Injection, Control, Staining

Journal: Frontiers in Pain Research

Article Title: Low-intensity muscle contraction exercise reduces pain sensitivity by modulating peripheral pathology and spinal sensitization in end-stage knee osteoarthritis rats

doi: 10.3389/fpain.2025.1644177

Figure Lengend Snippet: Immunohistochemical findings and quantification of immunoreactive cells for macrophage markers in the synovium. (A) Representative immunohistochemical findings show the CD68-, CD11c-, and CD206-positive cells in the synovium of the right knee joint at 56 days after MIA injection. Arrowhead: positive cells. Bar: 10 µm. (B) The number of CD68-positive cells. (C) The number of CD11c-positive cells. (D) The number of CD206-positive cells. *vs. the Sham group, #vs. the OA group, p < 0.05.

Article Snippet: The sections were blocked with 1% bovine serum albumin (BSA) containing 1% normal horse serum for 60 min. Next, the sections were incubated with a mouse anti-CD68 monoclonal antibody (Cat No. MCA341R, 1:200; Bio-Rad, Hercules, CA), rabbit anti-ITGAX/CD11c polyclonal antibody (Cat No. LS- B15988 , 1:2,000; LifeSpan Biosciences, Seattle, WA), and rabbit anti-mannose receptor polyclonal antibody (Cat No. ab64693, 1:3,000; Abcam, Cambridge, MA) for 30 min, followed by incubation with biotinylated horse anti-mouse IgG (BA-2000, 1:2,000; Vector Laboratories, Burlingame, CA) or biotinylated goat anti-rabbit IgG (H + L) (BA-1000; 1:3,000; Vector Laboratories).

Techniques: Immunohistochemical staining, Injection