i meos2 paxillin 22  (Addgene inc)

Journal: Bioactive Materials

Article Title: Pre-priming cell sheet therapy enabled by dynamic wrinkled electroactive substrate for muscle reconstruction

doi: 10.1016/j.bioactmat.2026.01.046

Figure Lengend Snippet: Schematic illustration of the NIR-responsive dynamic wrinkle platform for the non-invasive harvesting of pre-primed cell sheets and their application in volumetric muscle loss (VML) repair. (A) The process of obtaining and applying pre-conditioned cell sheets for VML repair. (B) NIR-triggered dynamic reconfiguration of the wrinkle topography remotely switches the interfacial adhesion state. (C) This reconfiguration alters cellular mechanotransduction and focal adhesion density, leading to cell sheet detachment when the interfacial mechanical force (Fm) surpasses the cell-substrate adhesion force (Fc). (D) Immunofluorescence staining of focal adhesion-related markers (integrin β1, talin, pFAK(Y397), paxillin, and YAP/TAZ) and cytoskeleton in cells under control and mechanical stimulation conditions.

Article Snippet: FAK Antibody (sc-271126), pFAK(Y3978556s), talin (sc: 4021s), paxillin (sc: 365379), integrin β1 (sc: 374429), and YAP (cst: 14074s), TAZ (cst: 83669s) were ordered from Santa Cruz Biotechnology.

Techniques: Immunofluorescence, Staining, Control

Journal: Bioactive Materials

Article Title: Pre-priming cell sheet therapy enabled by dynamic wrinkled electroactive substrate for muscle reconstruction

doi: 10.1016/j.bioactmat.2026.01.046

Figure Lengend Snippet: Immunofluorescence staining of focal adhesion-related markers and cytoskeleton in cells under control and mechanical stimulation conditions. Left (Control group): Immunofluorescence staining shows focal adhesion-associated proteins (green channels). These correspond to integrin β1(A), talin (B), pFAK/FAK (C), paxillin (D), YAP (E), and TAZ (F) in each row. F-actin and FAK are shown in the red channel. The rightmost column of each row displays merged images. These integrate focal adhesion marker signals (green), F-actin (red), and DAPI-stained nuclei (blue). Yellow indicates co-localization of focal adhesion markers and F-actin. Right (After mechanical stimulation group): Immunofluorescence staining displays the same set of focal adhesion-related proteins and F-actin in cells after mechanical stimulation. Merged images are presented in the same format as the control group.

Article Snippet: FAK Antibody (sc-271126), pFAK(Y3978556s), talin (sc: 4021s), paxillin (sc: 365379), integrin β1 (sc: 374429), and YAP (cst: 14074s), TAZ (cst: 83669s) were ordered from Santa Cruz Biotechnology.

Techniques: Immunofluorescence, Staining, Control, Marker

Journal: bioRxiv

Article Title: Quantitative multi-scale morphodynamic analysis reveals ratchet-like collective DVE migration and epiblast retrograde cell flow during anterior patterning in the mouse embryo

doi: 10.64898/2026.04.24.720339

Figure Lengend Snippet: (A) Diagram of E5.5 mouse embryo; Epi = epiblast, ExE = extra-embryonic ectoderm, EPC = ectoplacental cone, emVE = embryonic visceral endoderm, exVE = extra-embryonic visceral endoderm, DVE = distal visceral endoderm. (A’) Lateral view of example E5.5 Hex-GFP:mem-tdTomato embryo at two time-points, imaged using a ZEISS Z.1 light-sheet at a 10-minute interval. MIP = max intensity projection. Scale bar = 50 μm. (B) Polar geodesic projection of the apical VE surface of embryo in A’ . Re-projections enable visualisation of entire VE and the application of sophisticated motion analysis tools to analyse time-lapse data. Dotted line shows position of emVE-exVE boundary – the limit of DVE cell migration. Blue cross = distal tip. (C) Overview of VE motion-tracking pipeline for quantitative analysis of time-lapse data-set. (D’-D’’) Automatic staging of DVE migration from superpixel-based motion-tracking. (D) VE superpixel motion tracking on VE polar projection; red arrows show motion-flow vectors. DVE-core motion highlighted in green. Dotted line shows mean consensus angle vector of DVE which is used to align embryos along their A-P axis. The persistence of motion direction d direct of DVE was separated from the surrounding VE. (D’’) The motion persistence of DVE showed three phases; 1) pre-migration/initiation, 2) migration to boundary, 3) post-boundary, as shown in the max intensity projections. (D’’’) Example embryo showing last timepoint of each phase. (E) A flow-like pattern of motion across the VE was observed by superpixel motion tracking. Colours denote mean vector angle. (F) To track sub-regions of the VE, a deformable grid was seeded at the start of each phase. By tracking the motion within each sub-region the grid becomes deformed and multiple parameters are analysed. (F’) To quantify parameters and to combine data from across embryos, 2D tracking data is converted back to 3D coordinate-space for analysis which can be summarised in a multi-embryo polar plot. Note re-projections have no scale bar as they are non-linear.

Article Snippet: For Paxillin immunohistochemistry Hex-GFP stud males were crossed with CD1 females (Charles River) and dissected at E5.5.

Techniques: Migration, Plasmid Preparation

Journal: bioRxiv

Article Title: Quantitative multi-scale morphodynamic analysis reveals ratchet-like collective DVE migration and epiblast retrograde cell flow during anterior patterning in the mouse embryo

doi: 10.64898/2026.04.24.720339

Figure Lengend Snippet: (A) Example Hex-GFP; membrane td-tomato, (B) Lifeact-GFP, and (C) mTmG crossed with Ttr-Cre line. Each panel shows a max intensity projection, mid-sagittal optical section, and rectangular and polar projections of the apical surface of the VE (upper panels) and basal surface of the epiblast (lower panels). We note that the Lifeact-GFP mouse line consistently showed an almost complete absence of fluorescence signal in the ExE tissue. This does not reflect the level of F-actin in the ExE as shown by immunofluorescent staining. Note projections have no scale bar as they are non-linear.

Article Snippet: For Paxillin immunohistochemistry Hex-GFP stud males were crossed with CD1 females (Charles River) and dissected at E5.5.

Techniques: Membrane, Fluorescence, Staining

Journal: bioRxiv

Article Title: Quantitative multi-scale morphodynamic analysis reveals ratchet-like collective DVE migration and epiblast retrograde cell flow during anterior patterning in the mouse embryo

doi: 10.64898/2026.04.24.720339

Figure Lengend Snippet: (A) Example Hex-GFP; membrane td-tomato, (B) Lifeact-GFP, and (C) mTmG crossed with Ttr-Cre line. Each panel shows superpixel tracks on a rectangular geodesic projection and polar geodesic projections of the apical surface of the VE. Tracks are coloured by the direction of motion. Note re-projections have no scale bar as they are non-linear.

Article Snippet: For Paxillin immunohistochemistry Hex-GFP stud males were crossed with CD1 females (Charles River) and dissected at E5.5.

Techniques: Membrane

Journal: bioRxiv

Article Title: Quantitative multi-scale morphodynamic analysis reveals ratchet-like collective DVE migration and epiblast retrograde cell flow during anterior patterning in the mouse embryo

doi: 10.64898/2026.04.24.720339

Figure Lengend Snippet: (A) Overview of method for tracking the motion of VE sub-regions using superpixel motion tracking. (B) Example Hex-GFP;mem-tdTomato embryo showing the start, middle and end of migration in a geodesic polar projection. (C) Superpixel motion tracking of embryo in B. As the tissue moves the grid is deformed, enabling analysis of the tissue behaviour within each tissue sub-region and comparison across embryos. (D) Final timepoint of superpixel motion tracking from all (n = 21) embryos in this dataset. All data is converted back to 3D coordinates for analysis. For each stage (pre-migration/initiation, migration, post-boundary) the grid is re-set and re-tracked by motion behaviour. Note re-projections have no scale bar as they are non-linear. HXMT = Hex-GFP;membrane-tdTomato, LA = Lifeact-GFP, TTRmTMG = mTmG;Ttr-Cre embryo.

Article Snippet: For Paxillin immunohistochemistry Hex-GFP stud males were crossed with CD1 females (Charles River) and dissected at E5.5.

Techniques: Migration, Comparison, Membrane

Journal: bioRxiv

Article Title: Quantitative multi-scale morphodynamic analysis reveals ratchet-like collective DVE migration and epiblast retrograde cell flow during anterior patterning in the mouse embryo

doi: 10.64898/2026.04.24.720339

Figure Lengend Snippet: (A) Superpixel motion tracking of the emVE and exVE tissues confirms that emVE is significantly more active (higher mean speed and total speed) than the exVE (for both comparisons; Student’s t -test, p =<0.001). (B) Comparison of the anterior emVE and posterior emVE that are immediately adjacent to the DVE during the initiation of migration phase. There is no significant difference in motion (mean speed, total speed or anteriorwards velocity) or tissue area change (area fold change or area rate change) between these two tissue regions (all comparisons; Student’s t -test, p =>0.05). (C) Superpixel tracking of the distal VE during each phase shows that the migration phase has the highest anteriorwards velocity (one-way ANOVA, p =<0.01, followed by Tukey’s HSD Test on initiation of migration vs. migration (p =<0.001), migration vs post-boundary ( p =<0.001), initiation of migration vs post-boundary ( p =>0.05)). In the post-boundary phase the distal VE region contains the nascent Hex-GFP cells that continue to migrate anteriorly, but do so at a lower speed than the initial DVE.

Article Snippet: For Paxillin immunohistochemistry Hex-GFP stud males were crossed with CD1 females (Charles River) and dissected at E5.5.

Techniques: Comparison, Migration

Journal: bioRxiv

Article Title: Quantitative multi-scale morphodynamic analysis reveals ratchet-like collective DVE migration and epiblast retrograde cell flow during anterior patterning in the mouse embryo

doi: 10.64898/2026.04.24.720339

Figure Lengend Snippet: (A) Overview of cell movement analysis in cell tracked Lifeact-GFP time-lapse data-set, imaged at a 5-minute interval. Each cell vector angle is assessed then weighted by its velocity. (A’) Example embryo showing DVE cell tracks (cyan) on 2D polar geodesic projection. Red dotted line denotes emVE-exVE boundary. (A’’) Heat-map of weighted angle histogram of DVE cell velocity where each cell’s contribution is weighted by its speed. Cell vectors are in 22.5° bins for the +/-1 hour from onset of collective migration. Note the increase in coordinated anteriorward motion at the onset of the migration phase (white dashed line). (B) Graph showing Rayleigh distribution statistic of DVE cell tracks at pre-migration/initiation and migration phases, which shows the increase in coordination in cell vectors at the onset of migration (0 = random distribution, 1 = uniform distribution). (C) 2D geodesic polar projection of an example E5.5 Hex-GFP:mem-tdTomato embryo imaged every 10 minutes for 11 hours. Digitally extracting the A-P midline (coloured vertical line in C) throughout the time-lapse enables a kymograph to be plotted. (C’) Kymograph of the Hex-GFP channel along A-P mid-line of the embryo in C, showing the DVE cell population migrating collectively at the onset of the migration phase (vertical dotted line) to reach emVE-exVE boundary (horizontal dotted line). (C’’) Kymograph of superpixel motion tracking of A-P mid-line of embryo in C. Plotting anteriorwards velocity shows that the DVE movement is intermittent, undergoing start-stop behaviour (arrows). (D-D’) Graphs showing anteriorwards velocity and cumulative velocity (i.e., anteriorwards displacement/ min) of Hex-GFP population of embryo in C. In the pre-migration/initiation phase, where there is a relatively low level of directionally coordinated motion amongst DVE cells (A’’,B), there is little anteriorwards velocity. In the migration phase when directional coordination increased (A’’,B), anteriorwards velocity increases as the DVE migrated collectively (see position of Hex-GFP in D’’. t-5h - t0h). (E) Histogram of anteriorwards velocity at all timepoints from 9 Hex-GFP embryos during migration phase. In >90% of time-points Hex-GFP progresses anteriorly, with minimal posteriorwards motion. (E’) Plot showing mean time between start-stop events for each Hex-GFP embryo (n = 9); mean ratchet period was 1.13 ± 0.29 hours (mean ± stdev).

Article Snippet: For Paxillin immunohistochemistry Hex-GFP stud males were crossed with CD1 females (Charles River) and dissected at E5.5.

Techniques: Plasmid Preparation, Migration

Journal: bioRxiv

Article Title: Quantitative multi-scale morphodynamic analysis reveals ratchet-like collective DVE migration and epiblast retrograde cell flow during anterior patterning in the mouse embryo

doi: 10.64898/2026.04.24.720339

Figure Lengend Snippet: (A) Whole-mount immunofluorescence of PAXILLIN in an example E5.5 embryo, showing a maximum intensity projection (MIP) and single z-section of the embryonic-half of the embryo. PAXILLIN is enriched along the basal surface of the VE and epiblast. (B) Diagram of Cre-inducible EGFP-PAXILLIN reporter line. Ttr-Cre is expressed specifically in the VE and Sox2-Cre in the epiblast. (C) Example live imaging of E5.5 Ttr-Cre EGFP-PAXILLIN embryo and (D) Sox2-Cre EGFP-PAXILLIN embryo, showing the basal localisation of the protein throughout the VE and epiblast. A total of n = 15 embryos were imaged for both crosses. All scale bars 25 μm.

Article Snippet: For Paxillin immunohistochemistry Hex-GFP stud males were crossed with CD1 females (Charles River) and dissected at E5.5.

Techniques: Immunofluorescence, Imaging