Journal: Materials today. Bio

Article Title: Precise delivery of doxorubicin and imiquimod through pH-responsive tumor microenvironment-active targeting micelles for chemo- and immunotherapy.

doi: 10.1016/j.mtbio.2022.100482

Figure Lengend Snippet: Fig. 10. Immunostaining of tumor tissues in tumor-bearing mice after treatments. (A) Immunohistochemistry images of tumor sections stained with CD3, CD8, and TNF-α antibodies. The scale bar is 100 μm. (B) Immunofluorescence images of tumor tissues after treatments for 12 days. The scale bar is 50 μm. Blue fluorescence represents the cell nucleus stained with DAPI. Green fluorescence represents the iNOS stained with the iNOS antibody conjugated with FITC.

Article Snippet: After 30 min, the tissue slice was stained with diluted iNOS antibody (Miltenyi Biotec, catalog: 130-116-357) at 4 C overnight.

Techniques: Immunostaining, Immunohistochemistry, Staining

Journal: Biology of Sex Differences

Article Title: Sex differences in disease severity and immune responses in murine and human inflammatory arthritis

doi: 10.1186/s13293-026-00840-w

Figure Lengend Snippet: Male CIA mice exhibit higher disease severity compared to females. Male and female CIA and saline control mice were monitored for disease severity and assigned clinical scores from day 21 after the first CII challenge until the end of experiment (day 29). (A) Line graphs representing the mean clinical scores of mice starting from day 1 to day 29. On day 29 after the first CII challenge, mice were euthanized by cardiac puncture under anesthesia for blood and serum collection and storage at −80 °C. Serum concentrations of (B) anti-mouse collagen type II antibodies (left panel) and anti-bovine collagen type II antibodies (right panel) were analyzed by ELISA. N = 5 mice per group. One of two independent experiments. Simple linear regression analysis was performed to determine the statistical difference between the lines. One-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison post hoc test was used to determine the statistical significance between the groups. (*/ # p ≤ 0.05, ** p ≤ 0.005 and ***/ ### p ≤ 0.0005). In Fig. 1A, # significance of comparisons between CIA and saline control mice; *significance of comparisons between sexes of CIA mice

Article Snippet: Serum levels of mouse anti-collagen antibodies (autoantibodies) and bovine anti-collagen antibodies (antibodies to the immunizing antigen) were determined by ELISA using a Mouse Anti-mouse Type II Collagen IgG Antibody Assay Kit and Mouse Anti-Bovine Type II Collagen IgG Antibody Assay Kit, respectively, according to the manufacture’s protocol (Chondrex Inc. WA, USA).

Techniques: Saline, Control, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Neurochemical Research

Article Title: Morin Improves Cognitive Deficits in an in Vivo Model of Vascular Dementia by Modulating the N-methyl-D-aspartate Receptor Signaling Pathways

doi: 10.1007/s11064-026-04717-7

Figure Lengend Snippet: Morin modulated the expression of NMDA receptors in the hippocampus of VaD rats. a the expression levels of NR1 ; b the expression levels of NR2A ; c the expression levels of NR2B ; d the expression levels of NR1 protein; e the expression levels of NR2A protein; f the expression levels of NR2B protein; g protein levels of p-CREB; h protein levels of p-CAMK2A; i protein levels of p-CAMK2D. Protein levels of NR1, NR2A, and NR2B were quantified by ELISA. Data are presented as mean ± SD ( n = 8 per group). Statistical analysis was performed by one-way ANOVA with Tukey’s post-hoc test (data met assumptions of normality and homoscedasticity)/Kruskal-Wallis with Dunn’s test (data did not meet assumptions). * indicates a significant difference from the Sham group; # indicates a significant difference 2VO group; *# indicates a significant difference from both the Sham and 2VO groups, with a p-value of less than 0.05 considered statistically significant

Article Snippet: Moreover, phosphorylation levels of calcium/calmodulin-dependent protein kinase II isoforms CAMK2A and CAMK2D at Thr286 (p-CAMK2A, p-CAMK2D) were quantified using the Colorimetric Cell-Based ELISA Kit (CAMK2A/CAMK2D (Phospho-Thr286), Boster Bio, #EKC2366).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Materials Today Bio

Article Title: Neuro-bone-skin tri-regeneration via a microenvironment-responsive PRP-loaded chitosan hydrogel for traumatic brain injury therapy

doi: 10.1016/j.mtbio.2026.102913

Figure Lengend Snippet: CO@P hydrogel suppresses neuroinflammation. A-B) iNOS and Arg1 immunofluorescence: Day 3 vs. Day 28. Scale bar = 100 μm. C) Microglial polarization shift schematic (By Figdraw). D-E) Quantified fluorescence intensity of iNOS and Arg1 in each group. F-G) Quantitative analysis of the ELISA results of IL4, IL10, TNF-α and IL1-β on Day 3 and Day 28. Data were presented as mean ± SEM. N = 3 per group. ∗: P < 0.05 compared with Sham group; #: P < 0.05 compared with TBI group; Δ: P < 0.05 compared with CO@P group; .

Article Snippet: Primary antibodies used included NeuN (1100 μg/mL), GFAP (800 μg/mL), Iba1 (700 μg/mL), iNOS (820 μg/mL), Arg1 (900 μg/mL), VEGF (1000 μg/mL), CD31 (600 μg/mL), Caspase-3 (700 μg/mL), DCX (600 μg/mL) (1:200 dilution; Proteintech, China).

Techniques: Immunofluorescence, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Plasma Proteomics Identifies Thousand‐and‐One–Amino Acid Kinase 3 as a Potential Biomarker of Rheumatoid Arthritis Activity and a Novel Therapeutic Target

doi: 10.1002/art.70020

Figure Lengend Snippet: In vivo inhibition of TAOK3 alleviates bone destruction in CIA mice. (A) Schematic diagram of the CIA mouse experimental design. After a 7‐day acclimatization period, mice were immunized with bovine type II collagen for 28 days to establish the CIA model followed by intraperitoneal injection of sterile PBS (n = 6), MTX (n = 6), or SBI‐581 (n = 6) for 28 consecutive days. (B and C) Arthritis scores of hind paws and representative images from NI mice and CIA mice treated with different interventions. (D) Representative micro‐computed tomography images of hind paws and quantitative analysis of bone destruction scores across treatment groups. (E and F) Representative (E) H&E and (F) SO/FG‐stained sections from hind paw joints, with histologic scores for bone erosion, cartilage damage, and synovitis assessed according to Standardised Microscopic Arthritis Scoring of Histological sections guidelines (scale 0–3). Scale bar = 50 μm. In part B, data are presented as mean ± SD; in parts D–F, data are presented as individual values with group means indicated. Group comparisons were conducted using one‐way analysis of variance. * P < 0.05; ** P < 0.01; *** P < 0.001. CIA, collagen‐induced arthritis; H&E, hematoxylin and eosin; MMP, matrix metalloproteinase; MTX, methotrexate; NI, nonimmunized; PBS, phosphate‐buffered saline; SO/FG, safranin O/fast green; TAOK3, thousand‐and‐one–amino acid kinase 3. Color figure can be viewed in the online issue, which is available at http://onlinelibrary.wiley.com/doi/10.1002/art.70020/abstract .

Article Snippet: Twenty‐one days after the primary immunization, a booster injection was administered using bovine type II collagen emulsified with an equal volume of incomplete Freund adjuvant (Chondrex).

Techniques: In Vivo, Inhibition, Injection, Sterility, Micro-CT, Staining, Saline

Journal: Journal of Biological Chemistry

Article Title: Developmental Control of Apoptosis by the Immunophilin Aryl Hydrocarbon Receptor-interacting Protein (AIP) Involves Mitochondrial Import of the Survivin Protein

doi: 10.1074/jbc.m110.210120

Figure Lengend Snippet: FIGURE 6. Tom20 regulation of mitochondrial import of survivin. A, ali- quots of GST, GST-Tom20, or GST-Tom70 recombinant proteins were mixed with Raji cell extracts, and bead-bound material was analyzed by Western blotting. Bottom panel, Coomassie Blue staining of recombinant GST fusion proteins. B, Raji mitochondrial extract was immunoprecipitated with an anti- body to survivin or IgG, and proteins in pellets or supernatants (unbound) were analyzed by Western blotting. C, recombinant wild-type survivin or the survivin 1–141 mutant and GST-Tom20 beads were mixed with increasing concentration of recombinant AIP, and bead-bound proteins were analyzed by Western blotting (top panel). Bottom panel, GST or GST-Tom20 was incu- bated with recombinant AIP in the presence of increasing concentrations of survivin (up to 0.4 M), and bound proteins were analyzed by Western blot- ting. D, independently established clones (#) of HeLa cells stably transfected with control (Ctrl) or Tom70- or Tom20-directed shRNA were analyzed by Western blotting. E, HeLa cells were transfected with control, non-targeting (Ctrl) or Tom20-directed siRNA and analyzed by Western blotting. TCE, total cell extracts. F, clones of HeLa cells with stable knockdown of Tom70 or Tom20 were treated with 0.5 M staurosporine for 24 h and analyzed for cell viability by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide. DataaremeanS.E.ofreplicates(n3).***,p0.0001.Twodifferentclones per conditions were tested with identical results in two independent experiments.

Article Snippet: Antibodies against AIP (Novus Biologicals), survivin (Novus Biologicals), Tom20 (Santa Cruz Biotechnology), Tom70 (Novus Biologicals), COX-IV (Clontech), Smac (Pro-Sci), -actin (Sigma), and Hsp90 (BD Biosciences) were used.

Techniques: Recombinant, Western Blot, Staining, Immunoprecipitation, Mutagenesis, Concentration Assay, Clone Assay, Stable Transfection, Transfection, Control, shRNA, Knockdown