Journal: Journal of Cardiovascular Translational Research

Article Title: Extension of Atherosclerosis ApoE-/- Mouse—a Model of Chronic Myocardial Ischemia and Evaluation Method

doi: 10.1007/s12265-025-10734-8

Figure Lengend Snippet: A HE: It can be seen that the cardiomyocytes in the model group are significantly enlarged and the nuclei become sparse. Sirius red staining: Some areas of myocardium in the model group showed an increase in cell spacing accompanied by fibrotic tissue metaplasia. In contrast, myocytes in the control group were neatly arranged and densely distributed, with significantly less fibrous tissue than in the model group. “LV”: left ventricle. “RV”: right ventricle. B In the model group, a small number of apoptotic cells were observed with green fluorescence, but no large aggregates were detected. C Additionally, there were numerous CD68-positive cells expressing red fluorescence, which were irregularly distributed on the cell wall and in the cytoplasm

Article Snippet: Tissue sections underwent blocking with 5% bovine serum albumin (BSA) (Sigma-Aldrich Cat# A7906) in PBST (0.1% Tween-20 in PBS) for 1 h at room temperature, followed by overnight incubation at 4 °C with anti-CD68 primary antibody (Rabbit polyclonal, Bioss Antibodies, Cat# bs-20403R, RRID: AB_10855800) diluted 1:100 in blocking solution.

Techniques: Staining, Control, Fluorescence, Expressing

Journal: mBio

Article Title: Staphylococcus aureus exhibits spatiotemporal heterogeneity in Sae activity during kidney abscess development

doi: 10.1128/mbio.02043-25

Figure Lengend Snippet: Defining the four observed stages of S. aureus mouse kidney abscess development. C57BL/6 mice were inoculated RO with the S. aureus GFP − control strain. At the indicated time points (day 3 [D3] to day 5 [D5]), mice were sacrificed, and kidneys were harvested. ( A ) Representative images depicting stages of S. aureus kidney abscess development. Stage 1: intracellular S. aureus (white arrow), stage 2: small extracellular clusters (white arrow), stage 3: fully formed and intact SAC, and stage 4: dispersed SAC. ( B ) Areas of S. aureus extracellular clusters (stage 2) or SACs (stages 3 and 4) at D3 and D5. Dots represent individual extracellular clusters (stage 2) or SACs (stages 3 and 4). N = 3 to 4 mice per time point. Red bars: mean. ( C ) Immunofluorescence images showing S. aureus inside neutrophils (Ly6G + ). XZ and YZ planes corresponding to the point highlighted by the white arrow are shown at the top and right, respectively. ( D ) Immunofluorescence microscopy differentiating extracellular vs intracellular mCherry + S. aureus (within Ly6G + neutrophils) by staining with antisera (green) either before (top row) or after (bottom row) permeabilization. Extracellular bacteria were stained with antisera prior to permeabilization, while intracellular bacteria were only accessible to antisera after permeabilization. Dotted lines connect objects in the XZ and YZ planes. ( E ) Representative immunofluorescence images showing localization of neutrophils (Ly6G + , top row) and macrophages (CD68 + , bottom row) around a stage 3 SAC. Right column: zoom of the white boxed area. Statistics: ( C ) Kruskal-Wallis one-way analysis of variance (ANOVA) with Dunn’s post-test. * P < 0.05, n.s.: not significant.

Article Snippet: Sections were thawed in PBS at room temperature (RT) and stained with Hoechst (1:10,000 dilution in PBS) for 15 min. To stain immune cells, thawed sections were permeabilized using ice-cold methanol for 2 min, blocked with 2% bovine serum albumin (BSA) in PBS at RT for 1 h, then incubated with either APC-conjugated rat anti-mouse Ly6G antibody (Invitrogen: 17-9668-80) or rat anti-mouse CD68 primary antibody (BioRad: MCA1957T) diluted in 2% BSA overnight at 4°C.

Techniques: Control, Immunofluorescence, Microscopy, Staining, Bacteria

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