mouse antii 67 antibody  (Cell Signaling Technology Inc)

Journal: Developmental Neurobiology

Article Title: Molecular and Proteomic Profiling of Human Cerebral Organoid‑Derived Small Extracellular Vesicles

doi: 10.1002/dneu.70062

Figure Lengend Snippet: Effects sEVs obtained from static, orbital shaker and Organoid‐on‐a‐Chip conditions on U87 glioblastoma cells. (A) MTT assay for viability of U87 Glioblastoma for static, orbital shaker, and Organoid‐on‐a‐Chip sEVs. (B, C) Representative Ki67/DAPI immunofluorescence images of U87 glioblastoma cells following sEV treatment and quantification of Ki67–DAPI signal overlap using the Manders coefficient. (D, E) Migration of U87 Glioblastoma cells upon sEV treatment was observed by scratch assay up to 48 h. Scale bar 400 µm. (F, G) Diameter of spheres formed on day 2 and day 4 of sEVs treated U87 Glioblastoma by hanging drop sphere formation assay were quantified. Scale bar 200 µm. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01 compared to the control group.

Article Snippet: The primary antibody, Ki‐67 (Bioss, bs‐2130R), was added to the samples and incubated overnight at 4°C.

Techniques: MTT Assay, Immunofluorescence, Migration, Scratch Assay, Tube Formation Assay, Control

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Renal Tubular Epithelial Cells Senescence Induced by 1‐Aminopyrene Activates Fibroblast via BRD4 Driven Senescence‐Associated Secretory Phenotype Signaling

doi: 10.1002/jbt.71124

Figure Lengend Snippet: 1 ‐ AP induces tubular epithelial senescence and promotes paracrine fibroblast activation via a SASP‐mediated mechanism. (A) Representative images and quantification of SA‐β‐gal positive RTECs following exposure to 1‐AP after 72 h. Scale bars, 5 µm. (B) DAPI‐stained nuclei (blue) and quantification showing a significant reduction in cell number following exposure to 1‐AP after 72 h. Scale bars, 2 µm. (C) Representative images and quantification of immunofluorescence analysis of Ki67 in RTECs following 1‐AP treatment after 72 h. Scale bars, 5 µm. (D) RT‐qPCR analysis of p16, p21, and p53 mRNA expression showing upregulation of senescence‐associated cell cycle regulators in RTECs following exposure to 1‐AP after 48 h. (E) Representative images and quantification of immunofluorescence analysis of p21 in RTECs following 1‐AP treatment after 72 h. Scale bars, 5 µm. (F) Heatmap for RT‐qPCR analysis of senescence‐associated secretory phenotype (SASP) factors (IL‐1β, IL‐6, IL‐8, MCP‐1, TGF‐β1, and CXCL1) in RTECs following 1‐AP treatment after 48 h. (G) Schematic illustration of the experimental design showing 1‐AP induced senescence in RTECs and paracrine activation of renal fibroblasts via conditioned media may leads to TIF or renal fibrosis. (H) Heatmap for RT‐qPCR analysis of fibrotic markers ( α‐SMA, POSTN, Col1a2, TIMP1 , and CTGF ) in NRK52F renal fibroblasts exposed to conditioned media from 1‐AP‐treated TECs for 48 h. (I–K) Representative images and quantification of immunofluorescence analysis of α‐SMA (I), POSTN (J), and COL1A1 (K) in NRK52F fibroblasts, demonstrating significant fibroblast activation and extracellular matrix induction following exposure to TEC‐derived conditioned media after 48 h. Scale bars, 20 µm. Data are presented as mean ± SD, n = 3. Statistical analysis was performed using two‐tailed unpaired Student's t‐test. Exact p ‐values are indicated in the graphs; ns, not significant. DAPI used as nuclear counterstain.

Article Snippet: Primary antibodies for AhR antibody (182705‐10 G, Meck), Phospho‐Histone H2A.X (Ser139) Antibody (20E3, cell signaling), BRD4 antibody (ER1901‐02, Thermo‐Fischer), Ki‐67 antibody (27309‐1‐AP, proteintech), p21 (2947, cell signaling), α‐SMA antibody (14395‐1‐AP, proteintech), periostin (19899‐1‐AP, proteintech), COL1A1 antibody (55131‐1‐AP, proteintech), Histone H3K9/14ac (Acetyl H3K9/14) Antibody (A‐4021‐025, epigentek), Histone H3K27me3 (H3K27 Trimethyl) Antibody (A‐4039‐025, epigentek), Histone H3K9me3 (H3K9 Trimethyl) Antibody (A‐4036‐025, epigentek), IL‐6 (21865‐1‐AP, proteintech), TGF‐beta1 (21898‐1‐AP, proteintech) and Secondary antibody anti‐Mouse Alexa‐Fluro 555 ( A31570 , Thermo‐Fischer) and anti‐Rabbit Alexa‐Fluro 488 (A11008, Thermo‐Fischer), TUNEL Assay Kit (25879, cell signaling) and Senescence β‐Galactosidase (SA‐β‐gal) Staining Kit (9860, cell signaling) were purchased.

Techniques: Activation Assay, Staining, Immunofluorescence, Analysis, Quantitative RT-PCR, Expressing, Derivative Assay, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: Targeting the PCSK9–PR–IL-17 axis for psoriasis therapy

doi: 10.3389/fphar.2026.1884619

Figure Lengend Snippet: PR agonist alleviates psoriasis-like skin inflammation and suppresses IL-17 pathways. Male C57BL/6J mice (6–8 weeks old) were shaved on the dorsal skin 2–3 days before IMQ application to induce psoriasis-like dermatitis. Mice (n = 6 per group) were then treated with progesterone or vehicle for 4 days. (A) Changes in body weight during the experiment. Significance symbols shown below the curves indicate comparisons between the IMQ + Pg (0.3%) group and the IMQ group. (B) Representative macroscopic photographs and corresponding H&E-stained sections (bottom) of dorsal skin from the different treatment groups. (C) Quantification of psoriasis-like dermatitis severity (PASI-like score), including scores for erythema, scaling, and skin thickness. Significance symbols shown above the curves indicate comparisons between the IMQ + Pg (0.3%) group and the IMQ group, whereas symbols shown below the curves indicate comparisons between the IMQ + Pg (0.1%) group and the IMQ group. (D) Immunohistochemical staining for PCSK9, PCNA, Ki-67, and p-p65 in skin sections from each group (n = 3). (E) Immunofluorescence staining for CD11b and CD4 in skin sections from each group (n = 3). (F–I) Quantitative PCR analysis of mRNA levels ( CXCL1 , CXCL8 , S100A8 , and S100A9 ) in HaCaT cells treated with IL-17A and progesterone. Results were normalized to ACTB (n = 3). Data are presented as mean ± SEM; symbols represent individual animals or independent biological replicates. Statistical analyses were performed separately for each panel. For (B,D,E,F,G,H,I) , statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test. For (A–C) , longitudinal data were analyzed using repeated-measures two-way ANOVA with time and treatment as factors, followed by Tukey’s multiple comparisons test. Geisser–Greenhouse correction was applied when required. *P < 0.05, **P < 0.01, ***P < 0.001; ns, not significant.

Article Snippet: Anti-DYKDDDDK tag polyclonal antibody (20543-1-AP), phospho-NF-κB p65 (Ser536) recombinant monoclonal antibody (80379-2-RR), and Ki-67 polyclonal antibody (28074-1-AP) were obtained from Proteintech (United States).

Techniques: Staining, Immunohistochemical staining, Immunofluorescence, Real-time Polymerase Chain Reaction, Analysis