alexa fluor 488 conjugated rabbit monoclonal anti phospho histone h3 ph3  (Cell Signaling Technology Inc)

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: bioRxiv

Article Title: Live dynamics of induced cell-cell fusion between mitotic and interphasic cells

doi: 10.64898/2026.01.27.700572

Figure Lengend Snippet: (A) Comparison of Mad2 localization in control prometaphase and anaphase cells and mitotic fused cells undergoing premature mitotic exit. Mad2 can be detected at some kinetochores despite the presence of nuclear envelope membranes. (B) Time lapse of cerulean-Cyclin B1 transiently transfected in LLC-PK1 cells expressing H2B-mCherry after cell-cell fusion. Cyclin B1 was not degraded during induced mitotic exit of the mitotic cell. (C) Analysis of pH3-s10 in fixed prometaphase control cells and at different time points after cell-cell fusion. At 30 minutes after cell-cell fusion there is a mixed population of cells with high (second panel) and low (third panel) pH3-s10 levels, despite the presence of nuclear envelope membranes. At 60 minutes after cell-cell fusion, most cells show very low levels of pH3-s10. Magenta squares show magnification of nuclear envelope membranes around chromosomes. (D) Quantification of pH3-s10 in telophase control cells and at several time points after cell-cell fusion. Control, n=19 cells; 20 min, n=20 cells; 30 min, n=22 cells; 40 min, n=27 cells; 50 min, n=32 cells and 60 min, n=26 cells. Note that only 50 and 60 minutes after cell-cell fusion the levels of pH3-s10 become similar to control levels. Statistics, non-parametric Mann-Whitney test. Time is h:min. Scale bar is 10μm.

Article Snippet: Primary mouse mab414 (1:1000; ab24609, Abcam), mouse anti-α-Tubulin (1:500, DM1A, sigma), mouse anti-Mad2 (1:100, sc-65492, Santa Cruz) and rabbit anti-pH3-s10 (1:200, 3377, Cell Signaling).

Techniques: Comparison, Control, Transfection, Expressing, MANN-WHITNEY