Journal: Cell Death Discovery

Article Title: GPER1 reduces skin inflammation by inhibiting keratinocyte proliferation

doi: 10.1038/s41420-026-03059-1

Figure Lengend Snippet: a Schematic of the experimental procedure used. Spint1a-deficient embryos were treated with Palbociclib or vehicle (DMSO) at 1 dpf by bath immersion. At 2 dpf, images were taken to analyze Nfkb reporter expression and neutrophil distribution. Anti-pH3 immunofluorescence was also performed. b Representative images of 2 dpf control and treated larvae, showing neutrophils and Nfkb reporter activity. c Quantification of Nfkb activity in skin in each group. Quantification of distribution ( d ) and number ( e ) of neutrophils in tail in every condition. e Quantification of neutrophil distribution in the tail in every condition. f Representative images of anti-pH3 immunofluorescence of 2 dpf control and treated larvae. g Number of pH3 + cells in skin in every condition. h Analysis of the correlation between the number of neutrophils and the number of pH3 + cells in skin. Each dot represents one individual, and the means and SEM for each group are also shown. P values were calculated by Student’s t test ( c , d , f ) and by Pearson Correlation test ( g ). ns, not significant; * P ≤ 0.05 ; *** P ≤ 0.001.

Article Snippet: Following, they were incubated in anti-Phospho-Histone H3 (pH3) antibody (Cell Signaling, #9701, 1:100) diluted in blocking solution for two days at 4 °C.

Techniques: Expressing, Immunofluorescence, Control, Activity Assay

Journal: bioRxiv

Article Title: Single-Cell Cross-Species Profiling identifies Conserved Transcriptional Networks in Early Pancreatic Tumourigenesis

doi: 10.64898/2026.03.03.708839

Figure Lengend Snippet: (A) Schematic representation of the transgenes used to generate the control (Ac-G) and the acinar-derived pancreatic tumour model (Ac-K). (B-E) 5-day zebrafish larvae expressing the protein GFP ( B,D ) or GFP-KRAS G12D ( C,E ) in the acinar cells of the pancreas. White asterisk indicates gut autofluorescence. White arrows indicate the fluorescent heart, marked by the cmlc2:GFP reporter. Panels D and E show magnified views of the pancreata outlined with white dashed lines in panels B and C. i: principal islet, a: acinar cells. Scale bars : 200 µm. (F-G) Representative photographs of male zebrafish showing a marked abdominal protrusion, indicative of tumour presence ( G, white arrow), compared to a control male ( F ). Scale bars: 2mm. (H-I) Dissected digestive systems of an adult control zebrafish ( H ), showing a healthy pancreas (P, white dashes) or of an Ac-K p53 m/m fish ( I ) with a pancreatic tumour (T, yellow dashes). L: liver, G: gut, S: spleen, P: pancreas, T: tumour. Scale bars: 2mm. (J) Tumour incidence across various TP53 backgrounds. K-L: Hematoxylin and eosin (HE) staining of transversal sections of control pancreas (K) and Ac-K p53 m/m tumour (L) with close-up regions (K’,L’) outlined by black boxes. M-N : Immunofluorescence staining with phospho-histone H3 (pH3) antibody on control (M ) and tumoral tissue sections (N). Control and tumoral pancreatic tissues are surrounded by green dashes. L; Liver, G; gut, d; extrapancreatic duct, i; principal islet, a; acinar cells. Scale bars: 200 µm. O : Quantification of pH3-positive cells in four Ac-K p53 m/m tumours compared to six control pancreas (3 p53 +/+ ; 1 p53 +/m , 2 p53 m/m ). The percentage of pH3 positive cells was calculated as the ratio of the pH3-positive area to the DAPI-positive area, based on the mean of two independent sections per fish. Each point represents an individual fish. Data are presented as mean ± SD; **P = 0.0095, determined using the Mann–Whitney test.

Article Snippet: Immunofluorescence staining was performed with anti-GFP (rabbit, Cell Signalling #2956S 1:200), anti-Caveolin-1(rabbit, Cell Signalling #3238 1:250), anti-α-Amylase (rabbit, Cell Signalling #3796 1:200), anti-Phospho-Histone H3 (pH3) antibodies (rabbit, Cell Signalling #3377T 1:200) overnight at 4°C followed by rabbit HRP SignalStain Boost (Cell Signalling #8114), and Tyramide-FITC, Tyramide-Cy3 (TSA Plus Cyanine 3 System #NEL744001KT Akoya BioSciences) or Tyramide-Cy5 substrate (TSA Plus Cyanine 5 System #NEL745001KT Akoya BioSciences).

Techniques: Control, Derivative Assay, Expressing, Staining, Immunofluorescence, MANN-WHITNEY

Journal: Frontiers in Cell and Developmental Biology

Article Title: Pharmacological inhibition of frizzled 4 delays cell cycle progression and limits oral squamous cell carcinoma growth

doi: 10.3389/fcell.2026.1756565

Figure Lengend Snippet: Phenotypic characterization of cell cycle alterations following FzM1 treatment. (A) Representative immunofluorescence images of Ki67 (green) with nuclear counterstaining using DAPI (blue). Scale bar, 30 µm. (B) Quantification of the percentage of Ki67-positive cells relative to total nuclei. (C) Mitotic figures identified morphologically via DAPI staining. Scale bar, 30 µm. (D) Quantification of the percentage of cells in mitosis relative to total nuclei. (E) Representative immunofluorescence images of phospho-histone H3 (PH3, green) with DAPI (blue) nuclear counterstaining. Scale bar, 30 µm. (F) Quantification of the percentage of PH3-positive cells relative to total nuclei. (G) GSEA plots showing enrichment of pathways associated with specific cell cycle phases, mitotic structural changes, and key cell cycle regulators. (H) Representative images of EdU-positive cells over time during cumulative incorporation assays used to calculate cell cycle progression. Scale bar, 30 µm. (I) Graphical representation and quantification of the percentage of EdU-positive cells over time for cell cycle analyses. p-values for significant time point: 14 h, p = 0.024; 16 h, p = 0.00045; 20 h, p = 0.00421; 24 h, p = 0.03402. *p < 0.05; **p < 0.01; ***p < 0.001; not significant (p > 0.05). Data are presented as mean ± SEM.

Article Snippet: For immunofluorescence, cells were probed with anti-FZD4 (#SAB2108140; Sigma-Aldrich, 1:100), anti-phospho Histone3 (pH3) (#9701, Cell Signaling, 1:300); anti-Ki67 (#ab16667, abcam, 1:100) and detected with donkey anti-rabbit IgG Alexa Fluor 488 or 546 (1:1000; Invitrogen).

Techniques: Immunofluorescence, Staining