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: Nucleic Acids Research

Article Title: Phosphorylation of Barrier-to-Autointegration Factor (BAF) regulates anchoring of centromeric heterochromatin to the nuclear envelope (NE)

doi: 10.1093/nar/gkag021

Figure Lengend Snippet: Non-phosphorylatable BAF forms delay mitosis. ( A ) The proportion of G2/M cells determined by FACS is shown for S2 cells transiently expressing N-BAF, C-BAF, N-BAF T4A/S5A , or C-BAF T4E/S5E , and for control mock transfected S2 cells. Results are from a single experiment ( N = 3818 for N-BAF, 6565 for C-BAF, 6764 for N-BAF T4A/S5A , 818 for C-BAF T4E/S5E , and 59 729 for control; P -values: ns > .05; * < .05; **** < .0001; Chi-square test). ( B ) Immunostaining with αH3PS10 (in red) of cells transiently expressing N-BAF, C-BAF, or N-BAF T4A/S5A . N-BAF, C-BAF, and N-BAF T4A/S5A signals (in green) are direct fluorescence. DNA was stained with DAPI (in gray). Scale bars are 10 μm. ( C ) Quantification of the results shown in panel (B). The proportion of αH3PS10 positive cells is shown for S2 cells transiently expressing N-BAF, C-BAF, or N-BAF T4A/S5A , and for control mock transfected S2 cells. ( N = 389 for N-BAF, 275 for C-BAF, 235 for N-BAF T4A/S5A , and 748 for control; P -values with respect to control: **** < .0001; Fisher’s exact test).

Article Snippet: The rest of antibodies used are commercially available: mouse monoclonal αHP1a (DSHB C1A9), mouse monoclonal αLaminDm0 (DSHB ADL67.10), rabbit polyclonal αGFP (Invitrogen A11122), mouse polyclonal αGFP (Roche 118144600), rabbit polyclonal αFlag (Merck Life Science F7425), rabbit polyclonal αH3 (Cell Signaling 9715), rabbit polyclonal αH3K9me3 (Millipore 67–442), rabbit polyclonal αH3PS10 (Cell Signaling 9701), and mouse monoclonal αTubulin (MAB3408 Millipore).

Techniques: Expressing, Control, Transfection, Immunostaining, Fluorescence, Staining